OBJECTIVE: To explore the protocol that induced marrow mesenchymal stem cells (MSCs) differentiating into islel-secreting cells in vitro and to provide new clues for the sources of islet transplantation. METHODS: Using a defined culture medium and technique for transdifferentiation, MSCs from adult SI) rats were guided into specific insulin-secreting cells. The expressions of nestin and islet-specific hormones and proteins, such as insulin, glucagon, somatoslatin and pancreatic duodenal homeohox 1 (Pdx-1) were analyzed by indirect immunofluorescence cytochemistry staining before and after induction. The expressions of pancreatic islet cell differentiation-related transcripts, such as neslin, insulin 1, glucose transporter 2 (GLUT 2), Isl-f, Pdx-1, Pax-4 mid Pax-6 were detected by reverse transcription-PCR (RT-PCR). In addition, the quantity of insulin secretion was examined using radioinimuna-issay. RESULTS: Five hours after induction, (44.6 ± 7. 3)% of differentiated MSCs expressed nestin and it increased to (61. 8 ± 8. 4)% 24 hs after induction, but the expression of nestin almust disappeared at day 14. In the meantime, islet-like cellular clusters appeared after day 14 and became more apparent by day 28. Differentiated cells were found to be immunoreactive to insulin, glucagon, somalostatin and Pdx-1, and expressed insulin 1, GLUT 2, GK, Isl-1, PDX-1, Pax-4, Pax-6 mRNA. In addition, the results of cumulative quantities of insulin of 24 hs and the stimulation index showed that differentiated cells were able to produce insulin at higher levels, and displayed glucose-dependent insulin release in vitro. CONCLUSIONS: Adult rat MSCs can be differentiated into insulin-secreting cells in vitro. This approach might lead to widespread cell replacement therapy for Type 1 diabetes.

OBJECTIVE: To evaluate the value of acoustic densitometry (AD) in detecting myocardium damage of the left ventricle secondry to hypoxic pulmonary hypertension (PH). METHODS: Using the rabbit model of hypoxic PH, the density of various locations of the left ventricular myocardium in the hypoxic PH group and the normal control group were detected by the technique of AD. RESULTS: Compared with the normal control group, average image intensity (All) of the left ventricular anterior wall (LVA) in the hypoxic PH group increased ( P <0.05), while the difference between the standard deviation intensity (SDI) and peak to peak intensity (CVIB) of LVA were not significantly different ( P > 0.05). The CVIB of the middle of the interventricular septum (IVSM) in the hypoxic PH group was lower than that of the normal control group ( P <0.05). The All of IVSM and the left ventricle posterior wall (LVPW) in the hypoxic PH group were higher than those of the normal control group ( P < 0.05). In various locations of the left ventricular myocardium, the value of All and the left ventricle diastolic end pressure (LVDEP) were positively related ( r = 0.6206 -0.6311, P <0.01). CONCLUSIONS: The AD technique is useful in evaluating the extent of damage of the left ventricular myocardium in hypoxic PH.

OBJECTIVE: To study the changes of serum TNF-α and NO in neonatal rats with endotoxic shock and its relationship with myocardial cells damge and to explore the protection effect of dexamethasone (DXM) on neonatal rats with endotoxic shock. METHODS: Nine of the 117 seven-day-old healthy neonatal Wistar rats were used as the pre-experimental base value control group, and the other 108 rats were randomly divided into control group, endotoxic shock group (LPSgroup, LPS 5 nig/kg) and treatment group (DXM group, LPS 5 mg/kg + DXM 5 mg/kg). Before and after injection of LPS (2, 4, 6 and 24 hs), 9 rats in each group were sacrificed and blood samples were collected to detect TNF-a by ELISA and NO by nitrate reductase. Myocardial super-microstructure was observed under an electron microscope. RESULTS: ① In the LPS group, the concentration of serum TNF-α peaked at 2 h, and it decreased to the level of control group after 6 hs. In the DXM group, the level of TNF-α at 2 h was higher than that of control group ( P < 0.05), but lower than that of the LPS group ( P < 0.05). At 4 hs the level TNF-α in DXM group was lower than that of the LPS group ( P <0.05), but was not different than that of the control group ( P >0.05). From 6 h after injection, the differences of TNF-α levels in 3 groups were not obvious ( P >0.05). ② In the LPS group, the concentration of NO rose after 2 h ( P <0. 01), and peaked at 24 h ( P <0. 01). After 2 hs, the levels of NO in the DXM group were lower than those of the LPS group ( P < 0.01). ③ Six hours after the injection of LPS, a little mitochondria of myocardial cells in the LPS group appeared to develop vacuolae-like degeneration. At 24 h, most mitochondria of myocardial cells in the LPS group presententecl with vaculea-like degeneration and necrosis. The myocardial fibers were broken. While in the DXM group, the changes in the super-microstructure at 24 h were not as serious as those which talk place in the LPS group. CONCLUSIONS: TNF-α and No were involved in the damage of myocardial cells in neonatal rats with endotoxic shock. DXM could partly protect the myocardial cells from damage by inhibiting the production of TNF-α and NO.

OBJECTIVE: Hydrocephalus is one of the most common complications of neonatal intraventricular hemorrhage (IVH). In this study, the effect of serial lumbar puncture (LP) on severe neonatal IVH is evaluated. METHODS: The effects of serial LP on 30 neonates hospitalized with severe IVH at Xinhua hospital since 1993 was evaluated. Another 30 neonates with severe IVH who had not been treated for preventing post-IVH hydrocephalus were used as the control group. RESULTS: Of the 30 cases which received serial LP therapy, the cerebral ventricles of 25 cases stopped enlarging and shrank distinctly. The effective rate was 83.3% . The average age when serial LP therapy initiated was (15.0 ±13.5) days, the average treatment course was (16.9 ±12.9) days. The serial LP therapy was repeated (6.4±4.7) times. The avarage interval of LP was (2.9 ± 2.7) days and the average CSF volume removed was (6.7±1.6) ml. The average time of LP taking effect was (9.6 ± 5.5) days. Ten cases received diamox treatment during the course of serial LP. After 1 - 2 weeks' combined treatment, the enlarged ventricles of 8 cases returned to normal or remained steady. Eighteen of 25 ases were followed-up for (8.3±1.9) months. Their physical and mental development were normal. Cranialcerebral ultrasound examination found that the shapes of cerebral ventricles of 14 cases were normal and the cerebral ventricles of another 4 cases enlarged slightly. The treatment with removed CSF > 5 ml/ time or with a shorter interval of LP ( < 2 d) acheived better effect ( P < 0. 05 or 0. 01) . In the control group, the cerebral ventricles of 23 cases enlarged moderately or severely (6 of them complicated with hydrocephalus). CONCLUSIONS: Serial LP is a safe and effective method for severe neonatal IVH in preventing post-IVH hydrocephalus.

OBJECTIVE: Some patients who have been administrated benzodizepine for a long period will develop medicine tolerance. This study aims to investigate the molecular mechanism underlying this tolerance to benzodiazepine and the reversal of this tolerance by Rol5-4513. METHODS: One group of audiogenic seizure rats was administrated flurazepam for two weeks (the flurazepam group), which resulted in tolerance without behavioral signs of withdrawal to flurazepam. Another group was co-administrated Rol5-4513 daily for 7 days from the eighth day of flurazepam treatment (the Rol5-4513 group) to observe the effect of Rol5-4513 on the tolerance to flurazepam. The control group was administrated the same volume of propylene glycol as in the flurazepam group or the Rol5-4513 group. GABAA receptor subunit α1, α3, α5, γ2L and 72S were assayed using quantitative competitive RT-PCR in rat FrPaM and hippocampus. RESULTS: In the flurazepam group the content of mRNA encoding for α1, α3, γ2L and 72s was all significantly decreased (by 24%, 17%, 35% and 45% respectively) in FrPaM, whereas that of α5 was significantly increased (by 33%) compared with the control group. In hippocampus, α1, γ2L,and γ2S mRNA contents were significantly decreased (by 33% , 35% and 27% respectively). In the Rol5-4513 group, no significant changes were found with α1, α3,α5, γ2L and 72s in FrPaM, and α1,α5, γ2L, and γ23 in hippocampus compared with the control group. CONCLUSIONS: The accomodated change in GABAA receptor subunit α1 ,α3,α5, γ2L and γ2S in FrPaM and hippocampus may be associated with the mechanism for flurazepam tolerance in audiogenic seizure rats. Rol5-4513 can reverse the tolerance to flurazepam by affecting the modification of GABAA receptor subunit α1, α3, α5, γ2L, and γ2S subunit expression.

OBJECTIVE: It was thought that congenital heart disease (CHD) may be part of DiGeorge syndrome (DGS) and that the susceptibility to infection in children with CHD was related to immonodeficiency. This study explores whether immunodeficiency is present in children with CHD and aims to clarify the relationship between CHD and DGS by reviewing published papers. METHODS: The sizes of thymus shadow on chest X ray film were measured in 72 neonates with simple CHD, 34 neonates with complex CHD (n= 34) and 50 neonates with pneumonia. The partial immunologic laboratory data, including lymphocyte subsets counts, the peripheral blood monoeuclear cell (PBMC) IL-4 and IFN-7 mRNA expressions and production in culture supernatant, the PBMC proliferative response to phytahematoagglutinin (PHA) or lipopolysaccharide (LPS), and plasma IgG, IgA, IgM and complement 3 (C3) levels' were measured in 28 pre-school children with CHD and 20 age-matched healthy children. RESULTS: The thymus shadows on chest X-ray films were found in both neonates with CHD and neonates with pneumonia. There was no difference in the sizes of the thymus shadow between them. The peripheral lymphocyte subsets counts in children with CHD did not differ from those in the healthy children. There were also no differences in the plasma IgG, IgA, IgM and C3 levels between them. The counts of per minute impulse (cpm) of PBMC induced by PHA and LPS and the PBMC IL-4 and IFN-γ mRNA expressions in children with CHD did not differ from those in the healthy children. CONCLUSIONS: Not all children with CHD have congenital thymus aplasia or immunodeficiency. It is not certain that primary immunodeficiency is the reason why children with CHD may be prone to infection.

OBJECTIVE: To study the role of NF-κB signal pathway in neonatal rats with sepsis so as to provide the experimental base for corresponding clinical treatment of sepsis, in which NF-icB is taken as the target. METHODS: The sepsis model was established by injecting staphylococcus aureus subcutaneously in 10-day-old newborn rats. The activity of NF-icB in the lungs of newborn rats with staphylococcus aureus sepsis was detected by the electrophoretic mobility shift assay (EMSA) and the effect of anti-oxidant pyrrolidine dithiocarbamate (PDTC) on it was studied. RESULTS: In newborn rats with staphylococcus aureus sepsis, the activity of lung NF-κB was enhanced at the 1st h and reached a peak at the 3rd h after injection of staphylococcus aureus. PDTC had an inhibitive effect on the activity of lung NF-κB. The larger the dosage, the more intensified the inhibitive effect. CONCLUSIONS: In newborn rats with staphylococcus aureus sepsis, the NF-κB of lungs is activated, and the activation of NF-κB has a peak. The anti-oxidant PDTC can inhibit lung NF-κB activity in a dose-effect way in newborn rats with staphylococcus aureus sepsis.

OBJECTIVE: To evaluate the effect of anti-intercellular adhesion molecule-1 (ICAM-1) antibody on hypoxic-ischemic brain damage (HIBD) in neonatal rats. METHODS: Forty-eight SD neonatal rats were randomly assigned into four groups; HIBD group (Group A), anti-ICAM-1 antibody treatment HIBD group (Group B), normal saline treatment group (Group C) and normal control group (Group D). The rats in the Group A were sacrificed at 2, 24, 48, 72 and 168 hs respectively after resupply of oxygen, and the rats in the Group B and C were sacrificed at 48 h after treatment. The ICAM-1 expression of the brain was detected by immunohistochemical method at different time following the re-supply of oxygen. HE staining was used to observe neutrophil infiltration in the brain tissue and pathologic characteristics of brain cells. RESULTS: The expression of ICAM-1 was light at 2 h after the re-supply of oxygen and then increased at 24 h and reached its peak at 48 h in Group A. The neutrophil infiltration in the damage brain tissue of the Group A increased simultaneously. The expression of ICAM-1 and neutrophil infiltration in Group B significantly decreased compared with those of Group A at 48 h after the re-supply of oxygen. CONCLUSIONS: The expression of ICAM-1 may be correlated with the neutrophil infiltration in hypoxic-ischemic brain tissue. The anti-ICAM-1 antibody might have protective effects against HIBD in the neonatal rat.

OBJECTIVE: In recent years there has been an increasing interest in the evaluation of the neuronal responses to ischemic insult. Some cytokines, including transforming growth factor-beta 1 (TGF-β1) , are overexpressed after experimental brain demage in animals. This study aimed at the expression of transforming growth factor-betal (TGF-β1) in brain tissue after acute cerebral ischemia in mice. METHODS: Twenty mice were randomly assigned into a normal control group and a cerebral ischemia model group (n = 10 for each). The models of cerebral ischemia were established through ligating the left common carotid artery. The level of TGF-β1 mRNA expression was detected by in situ hybridization. RESULTS: In the model group, the brain tissue presented high levels of TGF-β1 mRNA expression: Grade Ⅲ in 8 cases and Grade Ⅱ in 2 cases, whereas in the normal control group only one case presented Grade Ⅰof TGF-β1 mRNA expression. CONCLUSIONS: TGF-β1 may participate in the pathogenesis of acute cerebral ischemia in mice.

OBJECTIVE: Energy failure of cerebral cells was regarded as one of the main mechanisms that led to hypoxic-ischemic encephalopathy(HIE) in the asphyxiated newborn. Few reports have discussed how to assess the relationship between energy failure and brain damage. The aim of the study is to explore the relationship between 3' , 5' -cyclin adenosine monophosphate (cAMP) and hypoxic brain damage by measuring the cAMP level in cerebrospinal fluid (CSF) of the asphyxiated newborn. METHODS: Thirty six full-term newborns were divided into three groups; moderate-severe HIE group (n= 12) , mild HIE group (n= 13) and non-HIE group (control group, n= 11). The levels of cAMP in CSF and plasma were measured by RIA between 36 hs and 72 hs after birth. The discharged newboms were followed up when they were 6 and 12 months old for measuring the mental development index (MDI) and psychomotor development index (PDI) by the Bayley Scales of Infant Development (BSID). RESULTS: The CSF cAMP level in the moderate-severe HIE group was (8.60±2.41) nmol/L, significantly lower than that of the mild HIE group [(14.83±2.84) nmol/L] and the control group [(24.43±2.39) nmol/L] (both P <0.01). There was a significant difference in the CSF cAMP level between the mild HIE group and control group ( P <0.01). The cAMP level in plasma in the moderate-severe group was the lowest, and that in the control group was the highest, but no significant difference was found between the mild HIE group and the control group. The results which were followed up showed that MDI and FDI of the moderate- severe HIE group were the lowest (84.79 ±13.34, 83.50±13.28, respectively); those of the mild HIE group were the second lowest (102.19±7.02, 99.94±9.08, respectively) and those of the control group were the highest (116.63±12.08, 116.69110.87, respectively). There were significant differences among the three groups ( P <0.01). ThecAMP level in CSF was significantly positively correlated to PDI and MDI ( r = 0.68, r =0.75,respectively, both P <0.01). CONCLUSIONS: There is a significant correlation between the cAMP level in CSF and hypoxic-ischamic brain damage. So the cAMP level in CSF following neonatal asphyxia might be used as one of the sensitive markers in evaluating the severity of brain damage at an early stage and predicting the future outcome.

OBJECTIVE: Low-molecular-weight heparins (LMWH) can inhibit proliferation of glomerular mesangial cells (GMCs) and it is a major and effective drug in the treatment of renal diseases but the mechanism has still not been explained. This paper studies the mechanism by which LMWH inhibits monocyte chemoaltractant protein-1 (MCP-1) expression, secretion and regulation in rat GMCs. METHODS: Three groups were established; GMCs group, LPS group (GMCs+ LPS) and LMWH group (GMCs + LPS + LMWH). GMCs proliferation was detected by MIT at 24 and 48 hs after culture. MCP-1 and nuclear transcriptional factor of Kappa B ( NF-κB) expressions were assayed by immunohistochemistry at 48 h after curture. MCP-1 concentration was determined by EL1SA. RESULTS: In the LMWH group with the terminal concentration of 250 IU/ml, the proliferative rate of GMCs was lower than that in the LPS group and was also lower than those in the LMWH groups with the terminal concentration of 2. 5 IU/ml and 25 IU/ml respectively ( P < 0.05). The proliferative inhibition of GMCs induced by LMWH of three different terminal concentrations was more significant at 48 h than at 24 h after culture. The positive rate of MCP-1 expression of GMCs in the LMWH group with the terminal concentration of 250 IU/ml was obviously lower than that in the LPS group at 48 h afterculture ( P <0.01), but there was no statistical difference when compared with the concentration in the GMCs group. The pasitive rale of MCP-1 expression of GMCs in the LPS group was obviously higher than that in the GMCs group ( P < 0.01). There was no statistical difference in the NF-κB expression at 48 h after culture between the LMWH group and LPS group as well as the GMCs group, while it was obviously higher in the LPS group than that in the GMCs group ( P < 0.01). The MCP-1 concentration in the LMWH group with the terminal concentration of 250 IU/ml was significantly lower than that in the LPS group ( P < 0.01) and did not differ from that in the GMCs group. CONCLUSIONS: LMWH can obviously inhibit proliferation of GMGs, down-regulate the abnormal expression and secretion of MCP-1, but can not inhibit the activity of NF-icB.

OBJECTIVE: To study the curative effect of two Chinese traditional medicines, astragalus membranaceus and folium isatidis, on murine viral myocarditis (VMC). METHODS: Balb/C mice were injected intraperiloneally with a diluted solution of coxsackievirus B3 (CVB3), and then randomly divided into three groups; infected control group, astragalus membranaceus treated group and folium isatidis treated group. The mice were sacrificed on days 3, 5, 7, 10, 14 and 21 after injection. The myocardial pathological score was estimated and viruses from myocardium were separated. In addition, the myocardial ultrastructure was detected under electon microscope on day 7 of post-injection. RESULTS: Compared with the myocardial pathological score of the infected control group, thxxse of both the astragalus membranaceus treated group (from day 5 to day 21) and folium isatidis treated group (on day 5 and day 7 post-injection) were significantly decreased ( P <0.05). There was no difference in the myocardial score after 10 days of infection between the folium isatidis group and the infected control group. The ultrathin section of myocardium in the infected control group revealed mitochondria! swelling, mitochondrial crista rupture and sarcoplasmic reliculum dilatation under electron-microscope. The pathological changes were relieved after treatment in both the astrgalus membranaceus and folium isatidis groups. Moreover, there were a lot of lysosomes in myocardium of the folium isatidis treated group. The results of viral isolation were positive on day 3 of post-infection in the three groups, with the same positive rate of 66. 7% . The same positive rate of viral isolation (100% ) was also found from day 5 to day 10 of post-infection in the three groups. On day 14 of post-infection, the positive viral isolation was noted only in the infected control group (33.3%) but failed in all three groups on day 21 of post-infection. CONCLUSIONS: Astragalus membranaceus may have a protective effect on myocardial cells. Folium isatidis could relieve myocardial lesions in the early stage of CVB3-induced myocarditis.

OBJECTIVE: The increase of oxygen-derived free radicals (OFR) or angiotensin Ⅱ(Ang Ⅱ ) in blood is one of the important causative agents of myocardium lesions. To develop a remedy for treatment of myocardium lesions induced by OFR or Ang Ⅱ , this study aims at exploring the protective effects of shenmai injection (SMI), growth hormone (GH), zinc sulfate (ZnSO4) and vitamin C (Vit C) on the cardiac myocytes injured by hydrogen peroxide (H2O2) or Ang Ⅱ. METHODS: H2O2 with the concentrations of 250 and 500 μmol/L or Ang Ⅱ with the concentrations of 10 6 and 10 7 mol/L were added respectively to the cardiac myocytes of neonatal rats so as to reduce the cardiac myocytes viability ( CMV). The injured cardica myocytes were administrated respectively with 5 ml/L and 10 ml/L (concentration) of SMI, 50 mg/L of Vit C, 15μmol/L and 60 μmol/L of ZnSO4, and 250 μmol/L and 500 μmol/L of GH. The increasing rate of CMV was measured at 24 h after medication. RESULTS: SMI, GH, ZnSO4 and VitC could all significantly elevate the increasing rate of CMV which had been decreased by H2O2 injury. The best effect was made by the SMI 10 ml/L group, followed in turn the ZnSO4 60 μmol/L, SMI 5 ml/L, Vit C 50 mg/L, GH 500 μmol/L, 15 μmol/L ZnSO4 and 250 μmol/L GH groups. There was no significant difference between the SMI 5 ml/L group and VitC 50 mg/L group. Nor was there among the GH 500 μmol/L, GH 250 μmol/L and ZnSO4 15 μmol/L groups. The CMV which had been decreased by Ang Ⅱ injury was significantly increased after administrating SMI and GH. The best effect was made by the SMI 10 ml/L group, followed in turn the SMI 5 ml/L, GH 500 μmol/L and GH 250 μmol/L groups. No difference was found among the last two groups. CONCLUSIONS: SMI, GH, ZnSO4 and VitC could all increase CMV which had been decreased by H2O2 injury, and both SMI and GH could increase CMV which had been decreased by Ang Ⅱ injury.

OBJECTIVE: Leptin, the obesity gene product, plays a key role in the regulation of energy metabolism and is correlated with fetal growth and development, but the mechanisms are not clearly understood. This study aimed to explore the relationship between soluble leptin receptor (sOB-R) in cord blood and fetal growth and development. METHODS: According to the birth weight, 67 full-term newborns were divided into two groups; the small for gestational age group (SGA group, n = 23) and the appropriate for gestational age group (AGA group, n = 44). The levels of leptin and sOR-R were assayed by ELISA and the nutrional state of the newborns was evaluated with body fat content measured by the Weststrate formula. RESULTS: The sOB-R level in cord blood was negatively related to leptin in cord blood, birth weight and body fat content ( r = -0.405, -0.366, -0.356 respectively; all P <0.05) and it was not related to maternal leptin level and sOB-R. The sOB-R level in cord blood in the SGA group was significantly higher than that in the AGA group [(18.24±6.02) ng/ml vs (13.8014.37) ng/ml; P <0.01], whereas the leptin level in the SGA group was much lower than that in the AGA group [(6.79±4.59) ng/ml vs (16.30±11.62) ng/ml; P<0.01]. The sOB-R level in cord blood in male infants was higher than that in female ones [(16.8914.37) ng/ml vs (13.9515.29) ng/ml; P <0.05], but the leptin level in the former was lower than that in the latter [(10.2818.28) ng/ml vs (15.70 ±12.11) ng/ml; P <0.05]. CONCLUSIONS: sOB-R in cord blood may have effects on fetal growth and development by regulating the serum level of free leptin. Quantification of both circulating leptin and sOB-R levels may be more valuable for understanding the mechanism of leptin.

OBJECTIVE: To study the clinical and epidemiologic features of burns in children and to take effective measures for preventing children from being burnt. METHODS: The clinical data of 367 cases of children burnt during 1994 - 2002 were studied retrospectively. RESULTS: Childhood burn cases accounted for 23.1% of the burn cases admitted to the Second Xiangya Hospital. The gender ratio of the childhood bum cases (boys/girls) was 2. 53. The incidence of burns in children from 1 to 3 years old was the highest (67.2%). The most common cause of burning in children from 1 to 3 years old and school children was hot water; for pre-school children it was fire. Severe burn injury, with the burn range of 15 % to 25 % of TBSA (total body surface area), was the most common (35.2 % ). CONCLUSIONS: Childhood burn has its own clinical and epidaniologic features. It is necessary to take effective measures to reduce the incidence of bums in children.