OBJECTIVE: Pulmonary hypertension is a proliferative vascular disease characterized by pulmonary vascular structural remodeling.Until now,the pathogenesis of pulmonary hypertension has not been fully understood. Recent many studies have suggested that endogenous vascular elastase plays a pivotal role in the vascular changes associated with pulmonary hypertension.The study aimed to explore the relationship between elastolytic activity and pulmonary remodeling in rats with pulmonary hypertension induced by monocrotaline. METHODS: One hundred mature Sprague-Dawley rats were randomly assigned into Normal and Model groups (both n=50). Rats in the Model group were induced with pulmonary hypertension by subcutaneous injection of monocrotaline (60 mg/kg). The elastolytic activity was measured by a fluorescence microplate reader and the pulmonary artery pressure was detected by catheterization at 2, 8,14 and 21 days post-injection. Light microscopic structural analysis of the peripheral pulmonary vasculature was performed at the same time points. RESULTS: The elastolytic activity increased significantly to(3.87±0.19)×10~(-8) U/mg at 2 days, and then decreased to (0.18±0.2)×10~(-8) U/mg at 8 days after injection, but on the 14th day it increased again to(1.45±(0.18)×)10~(-8) U/mg and remained high at (1.91±0.18)×10~(-8) U/mg on the 21st day.The extension of muscle into distal arteries was observed on the 8th day post-injection.Increased pulmonary artery pressure and medial wall thickness were present on the 14th day post-injection. CONCLUSIONS: The early increase in the elastolytic activity may be related to the initiation of pulmonary hypertension.The second increase may be involved in the development of this disease.
OBJECTIVE: Pulmonary surfactant protein-D(SP-D) is regarded as a valuable biomarker in acute lung injury(ALI) and acute respiratory distress syndrome (ARDS), but the alterations of SP-D in lung tissues in the early course of ALI remain unknown.This study was designed to explore the temporal fluctuations of SP-D and SP-D mRNA in young rats with ALI induced by lipopolysaccharide(LPS),as well as the alterations of ultrastructures of alveolar type Ⅱ(ATⅡ) cells. METHODS: Rat ALI models were established by intraperitoneal injection of LPS (4 mg/kg). The rats were sacrificed at 6,12, 24, 36, 48 and 72 hrs after LPS injection (8 rats each time point). Western blot and RT-PCR were employed to detect the contents of SP-D and SP-D mRNA in lung tissues. The ultrastructures of ATⅡcells were studied with transmission electron microscopy. RESULTS: Both SP-D mRNA and SP-D levels decreased after 12 hrs of LPS administration. The SP-D mRNA level reached a nadir at 24-36 hrs, but the SP-D level was reduced to its nadir by 48 hrs after LPS administration.LPS resulted in the alterations of lamellar bodies (LBs) in size (multilamellar forms), density(vacuole-like deformity) and number. The alterations of ultrastructures of ATⅡcells were most significant at 48 hrs. The clinical symptoms of ALI rats were most severe at 48 hrs. CONCLUSIONS: The alterations of the SP-D level were time-dependent in the early course of LPS-induced ALI. The lowest level of SP-D occurred at 48 hrs while severe multideformities of ATⅡcells were presented. A decreased level of SP-D in the lungs in the early stage of ALI may be associated with a worse clinical outcome.
OBJECTIVE: This study examined the prevalence of deletion and subtle mutations of survival motor neuron (SMN) gene in children with spinal muscular atrophy(SMA). METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the deletion of SMNt exon 7 in 25 children with SMA (type I 5 cases, type II 3 cases, and type III 17 cases) and in 24 healthy relatives of these patients. SMA was diagnosed clinically and pathologically. The subtle mutations of SMN in encode regions were screened by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with DNA direct sequencing in the patients without SMNt deletion and their relatives. RESULTS: Deletion of exon 7 of the SMNt gene was found in 5 cases of SMA type I(100%), 3 cases of type II (100%) and 6, type III(35%). No subtle mutation of SMN was found in encoded regions in 11 cases of type III SMA without SMNt deletion. The 24 relatives of SMA patients did not show the deletion and subtle mutation of SMN. CONCLUSIONS: ①Detection of SMNt gene exon 7 deletion can be recommend as a definitive diagnostic method for SMA, and showed promise to replace invasive examinations, such as electromyogram and muscular biopsy. ②In patients with type III SMA without SMNt deletion, diagnosis is still made clinically. ③No subtle mutation of SMN was found in SMA type III patients without SMNt deletion, suggesting genetic heterogenicity might exist.
OBJECTIVE: Interleukin 6(IL-6) is an autocrine growth factor for mesangial cells.This study aimed to investigate the changes of IL-6 levels in serum and urine of children with primary nephrotic syndrome (PNS) before and after glucocorticoid treatment. METHODS: Thirty-eight children with PNS, including 30 cases with steroid-responsive and 8, steroid-resistant,and 19 age-matched healthy children were enrolled in this study. Serum and urinary IL-6 levels were examined by ELISA before and after prednisone treatment (2 mg/kg daily for 8 weeks). RESULTS: Before prednisone treatment, the serum levels of IL-6 in the steroid-responsive group (118.74±31.18 ng/L) and steroid-resistant group (129.62±28.14 ng/L) were significantly higher than those in the normal controls (35.13±16.21 ng/L)(P<0.05). The IL-6 levels in urine of the steroid-responsive and steroid-resistant groups (14.19±4.87 and 22.54±5.35 ng/L) were also significantly higher than those in the normal controls(3.62±1.87 ng/L) (P<0.05). The IL6 levels of urine were significantly different between the steroid-responsive and steroid-resistant groups.After the treatment with prednisone for 8 weeks, the levels of IL-6 in serum and urine in the steroid-responsive group were significantly reduced to 41.68±18.94 and 5.11±1.31 ng/L respectively, which were not different from those in the normal controls. But the IL-6 levels in serum and urine of the steroid-resistant group did not decrease significantly after prednisone treatment. CONCLUSIONS: The detection of IL-6 levels in serum and urine is useful for the identification of steroid-responsive PNS and for the estimation of the prognosis of PNS.
OBJECTIVE: The treatment of steroid-dependent nephrotic syndrome (SDNS) remains a challenge.This study evaluated the efficacy of vincristine(VCR) in children with SDNS who had relapses after cyclophosphamide treatment. METHODS: The clinical data of 14 children with SDNS between June 2000 and March 2003 were studied retrospectively. They had relapses after more than one course of cyclophosphamide treatment before VCR treatment.VCR was administered by intravenous boluses in a dose of (1.0-1.5) mg/m~2 once weekly for 4 weeks then once monthly for 4 months. RESULTS: Thirteen patients completed the full course of VCR treatment and one patient discontinued after 6 times of VCR injection because of frequent relapses during the VCR treatment.Six of the 8 patients in relapse achieved complete remission after 2-3 times of VCR injection. After completing the full course of VCR treatment,12 patients achieved remission. Four remained in remission for the median time of 13.5 months (range 9-40 months). Seven of the 8 patients who continued to relapse received another VCR treatment (1 mg/m~2). After 1 or 2 infusions of VCR their proteinuria disappeared. Compared with before VCR treatment, the relapse frequency in half a year post-VCR treatment was decreased from 1.67 to 0.67 time (P<0.05). Side effects related to VCR treatment were uncommon. Only 3 patients presented with abdominal pain, but the symptom was alleviated after the dosage was reduced from 1.5 mg/m~2 to 1.0 mg/m~2. CONCLUSIONS: VCR treatment can induce complete remission of relapse and decrease the relapse frequency in children with SDNS.A few additional doses of VCR infusions may be effective in patients after a full course of VCR treatment but continue to relapse later.
OBJECTIVE: This study aimed to explore the relationship of the cellular immune function and the development of hypoxicischemic encephalopathy(HIE) by examining the levels of T subsets, membrane interleukin-2 receptor(mIL-2R), IL-1β, IL-6 and IL-10 in the peripheral blood of neonates with HIE. METHODS: The subjects included 32 term neonates with HIE and 30 healthy term neonates. The levels of CD3~+, CD4~+, and CD8~+, the ratio of CD4~+/CD8~+ and the mIL-2R levels before and after phytohemagglutinin (PHA) inducement were detected by biotin-streptavidin (BSA) on days 1, 3, and 7 after birth. Meanwhile, the levels of IL-1β, IL-6, and IL10 in serum were detected using ELISA. RESULTS: On the 1st day after birth,the levels of CD3~+, CD4~+ and CD8~+ and the ratio of CD4~+/CD8~+ in neonates with HIE were significantly lower than those in the normal controls. The mIL-2R levels before or after PHA inducement in HIE neonates were also significantly lower than those of the normal controls. On the 3rd day after birth, the levels of CD3~+, CD4~+ and CD8~+, and the ratio of CD4~+/CD8~+ in neonates with HIE remained lower than those in the normal controls. The mIL-2R level increased after PHA inducement in HIE neonates, but was still lower than that in normal controls. On the 7th day after birth, the levels of CD3~+, CD4~+ and CD8~+ in HIE neonates still remained lower than those in normal controls, but the ratio of CD4~+/CD8~+ and the mIL-2R level after PHA inducement were similar to those of the normal controls. The changes of T subsets and mIL-2R levels were most marked in neonates with severe HIE, followed by in those with moderate HIE. The serum levels of IL-1β,IL-6 and IL-10 in neonates with HIE increased significantly compared with those in normal controls on the days 1, 3, and 7 after birth. The increase was associated with the severity of HIE. CONCLUSIONS: Cellular immune function disturbance may exist in neonates with HIE. The expressive levels of cellular immune can be used as markers for early diagnosis of HIE and the evaluation of the severity of brain injury.
OBJECTIVE: There is controversy as to whether vitamin E supplementation is essential for healthy breast-feeding premature infants. This study examined the dynamic changes of the serum vitamin E level in healthy breast-feeding premature infants within 30 days after birth. METHODS: Thirty-eighty healthy breast-feeding premature infants were randomized to receive vitamin E supplementation (Intervention group, n=20) or without (Control group, n=18). The Intervention group was intramuscularly injected with vitamin E 50 mg daily for 3 days soon after birth.Blood samples were collected at birth (before the first vitamin E injection), and 10 and 30 days after birth. The serum vitamin E level was measured by spectrophotofluorometer. RESULTS: The vitamin E level of the Intervention group (1.64±0.68 mg/dL) was no significantly different from that of the Control group (1.76±0.74 mg/dL) (P>0.05) at birth. The Vitamin E level of the both groups increased 10 days after birth, and there was no significant difference between the two groups (2.54±1.23 mg/dL vs 2.64±1.13 mg/dl,P>0.05). On the 30th day after birth, the vitamin E level of the Intervention group remained high but that of the Control group decreased, although there was no significant difference between the two groups (2.77±1.56 mg/dL vs 2.37±1.07 mg/dL, P>0.05). No patient had the serum level of less than 0.5 mg/dL at any time point (reference range (0.59-6.45) mg/dL). CONCLUSIONS: There was no significant difference in serum vitamin E level between the two groups within 30 days after birth.The healthy breast-feeding premature infants were not vitamin E deficient.Thus Vitamin E supplementation is not necessary for healthy breast-feeding premature infants.
OBJECTIVE: Neonatal respiratory distress syndrome (RDS)is an acute clinical neonatal disorder that requires prompt management. This study aimed to evaluate the usefulness of the stable microbubble test (SMT) on gastric aspirates in the early diagnosis of RDS in premature infants. METHODS: One hundred and one samples of gastric aspirates obtained within half an hr of delivery from premature neonates born at the Iwate Medical University, Japan, with gestational ages between 24 and 35 weeks (mean 30.1 ±3.5 weeks) were detected by the SMT. The stable microbubles per mm~2 were counted. Chest X-rays were taken within 1 hr of delivery. The clinical characteristics and X-ray results were used as the gold standard for the diagnosis of RDS. The sensitivity,specificity and positive and negative values of the SMT to predict and make an early diagnosis of RDS were evaluated. RESULTS: Of the 101 premature infants, 31 were diagnosed as RDS by the gold standard.Twenty-two infants had stable microbubbles less than 10 bubbles/mm~2,7 had stable microbubbles between 10 and 20 bubbles/mm~2,and 2 had stable microbubbles more than 20 bubbles/mm~2.In the 70 non-RDS infants,only 1 had stable microbubbles less than 10 bubbles/mm~2,2 had stable microbubbles between 10 and 20 bubbles/mm~2,and the rest had stable microbubbles more than 20 bubbles/mm~2.The sensitivity and specificity of the SMT with a cut-off value of less than 10 bubbles/mm~2 to predict RDS were 70.97% and 98.57%,respectively,with positive and negative predictive values of 95.65% and 88.46%,respectively.With a cutoff value of less than 20 bubbles/mm~2,the sensitivity,specificity and positive and negative values were 93.55%,95.71%,90.63% and 97.10% respectively. The patients with bubbles less than 10/mm~2 (whose chest X-ray showed moderate RDS) were administered with surfactant replacement.As a result the patients' symptoms were significantly improved. CONCLUSIONS: The SMT on gastric aspirates is a rapid, simple and reliable procedure to predict and make an early diagnosis of neonatal RDS. It is helpful to the prophylactic exogenous surfactant replacement.
OBJECTIVE: In order to study the mechanisms of cerebral energy failure after hypoxic-ischemic brain damage (HIBD), the effects of hypoxia-ischemia (HI) on glucose transporter proteins (GLUT1 and GLUT3) expression in the brain were investigated in neonatal rats. METHODS: Thirty seven-day-old Wistar rats were randomly assigned into Normal (n=5), Sham-operated (n=5) and HIBD model groups (n=20). The HIBD model was established by ligating the right common carotid artery combined with the hypoxia exposure (8% oxygen). The rats were sacrificed at 1,3,5 and 10 days after HIBD. The expressions of GLUT1 and GLUT3 in the brain were examined by immunohistochemistry. RESULTS: Microvascular GLUT1 was seen in the Normal group. GLUT1 immunoreactivity began to increase in ipsilateral hemisphere 1 day after HI,peaked on the 3rd day, and remained high on the 5th day. No significant changes in GLUT3 immunoreactivity were observed 1 day after HI. Three days after HI, there was a pronounced decrease in GLUT3 staining; and on the 5th day the decrease of GLUT3 staining was most significantly. The maximal decrease of GLUT3 staining was seen in the hippocampal CA1 region. CONCLUSIONS: HI may result in an abnormal expression of glucose transporter proteins in the brain, which might aggravate the neuronal injury and interfere with its repair.
OBJECTIVE: Some studies have found that both heme oxygenase (HO) /carbon monoxide (CO) system and γ-aminobutyric acid B receptor subunits are involved in the pathogenesis of febrile seizure (FS). The aim of the study was to explore the effect of HO/CO system on GABA_BR subunits during FS. METHODS: Thirty-two Sprague-Dawley rats aged 21 days were randomly assigned into four groups: Control (37.0℃ water), FS, FS+hemin and FS+ZnPPⅨ groups (n=8 each). FS was induced in the last three groups by exposure to a hot water bath (45.0℃ water).Hemin(50 mg/ kg) and ZnPPⅨ(45 μmol/ kg) were intraperitoneally injected half an hour after the water bath in the FS+hemin and FS+ZnPPⅨ groups respectively. The Control and FS groups were administered with normal saline instead. The plasma level of CO was detected by the dual wavelengh spectrophotometer. The expressions of GABA_BR subunit mRNA and c-fos gene were examined by in situ hybridization. The expressions of GABA_BR subunit and Fos protein were observed by immunohistochemistry. RESULTS: The CO content in the FS group was significantly higher than that of the Control group. Hemin treatment increased the CO content while ZnPPⅨ treatment decreased the CO content.The expressions of GABA_BR_2 and GABA_BR_1 increased in the FS+hemin group compared with those in the FS group. In the FS+ZnPPⅨ group, the expression of GABA_BR_2 decreased while the GABA_BR_1 expression was not different compared with that in the FS group.Hemin treatment decreased the expression of c-fos gene and Fos protein, while ZnPPⅨ treatment elevated their expression. CONCLUSIONS: CO may play a role in the pathogenesis of FS through regulating GABA_BR function.
OBJECTIVE: This study examined the changes of neural stem cells (NSCs) following hypoxic-ischemic brain damage (HIBD) in order to provide a basis for clinical use of NSCs. METHODS: Neonatal 7-day-old rats were randomly assigned into three groups: Normal control, Hypoxic and Hypoxic-ischemic groups. The Hypoxic-ischemic group was subjected to the left common carotid artery ligation followed by 8% oxygen exposure for 2.5 hrs.The Hypoxic group was exposed to 8% oxygen for 2.5 hrs but without the carotid artery ligation.The subjects were sacrificed at 3 and 6 hrs, and 1, 3, 7, 14 and 21days after hypoxia/ischemia (10 rats at each time point). The number of NSCs from brain tissues of rats was detected with hematoxylin-eosin staining and immunohistochemistry. RESULTS: NSCs were presented in the normal brain tissues of neonatal rats and were reduced at 10 days postnatal.The NSCs in the Hypoxic and the Hypoxic-ischemic groups increased after hypoxia/ischemia and were significantly higher than that in the Normal control group at 6 hrs after hypoxia and at 1 day after hypoxia-ischemia. The NSCs number peaked at 3 days after hypoxia/ischemia and then decreased gradually but remained higher than that in the Normal control group at 21 days after hypoxia. CONCLUSIONS: NSCs may proliferate in the early phase of HIBD but decrease while the damage is developing. Early NSCs intervention for the treatment of HIBD appears to be promising.
OBJECTIVE: S100A4 gene plays an important role in neuroblastoma cell invasion and metastasis. This paper aimed to evaluate the liposome-mediated transfection efficacy of S100A4 antisense oligodeoxynucleotide (AS-ODN) and its stability in neuroblastoma cells. METHODS: The fluorescence (FAM) labeled S100A4 AS-ODN was transfected with Lipofectamine~(TM) 2000 into human neuroblastoma cells (LA-N-5). The transfection efficacy, stability and persistence time were observed by laser scanning confocal microscopy, and were compared with those of naked AS-ODN without Lipofectamine~(TM) 2000. RESULTS: The transfection efficacy of the S100A4 AS-ODN mediated by Lipofectamine~(TM) 2000 in neuroblastoma cells was high.The maximal signal intensity of intracellular fluorescence was 173 at 8 hrs after transfection and remained as high as 135 at 24 hrs. The maximal efficacy of transfection was 68%, and the stable expression lasted for more than 72 hrs. The transfection efficacy of the naked AS-ODN without Lipofectamine~(TM) 2000 was 35%. The maximal signal intensity of intracellular fluorescence was 90 and occurred at 6 hrs after transfection. The stable expression lasted for only 12 hrs. CONCLUSIONS: The S100A4 AS-ODN can be effectively transfected with Lipofectamine~(TM) 2000 into neuroblastoma cells, and be stably expressed,which may be useful in gene therapy for neuroblastoma
OBJECTIVE: Salviae miltiorrhizae (RSM), a Chinese medicine known as Dan-shen with the effect of promoting blood flow,has been used as a treatment for cardiovascular and cerebrovascular diseases. This study aimed to investigate the prophylactic effect of RSM on acute gastric mucosal injury in young rats. METHODS: Gastric stress ulcer were induced by water immersion restraint stress in 30 Sprague-Dawley young rats. The 30 rats were randomly assigned into Control and Experimental groups. The Experimental group was further subdivided into Normal saline and RSM groups according to the different medicines injected before stress. The rats of the RSM group were randomly administered 2 times, 4 times or 6 times by intraperitoneal injection of RSM, 2 mL each time 8 hourly. The Control group received no treatment. The histological changes of gastric mucosa and gastric mucosal ulcer index (UI) were observed. RESULTS: There was no difference in the UI between the Normal saline and Control groups.RSM pretreatment alleviated the gastric mucosal injuries. The rats that received 4 or 6 times RSM injection had significantly lower UI than the Control group. CONCLUSIONS: RSM has protective effects against gastric stress ulcer in young rats.
OBJECTIVE: To establish a relatively simple,reliable and reproducible in vitro model of the blood brain barrier (BBB). METHODS: Cortical brain microvascular endothelial cells (BMEC) and astrocytes (AS) from 7-day-old rats were co-cultured in cell culture inserts.VIII-factor and GFAP immunocytochemistry were used to identify the cell types of the BBB model. Cell morphological changes were observed under light and transmission electron microscopes.At the same time, the BBB restrictive characteristics were assessed by ~(125) I-BSA permeability. RESULTS: The positive rates of BMEC and AS were 95 % and 99%respectively. The densely packed cells were observed and the characteristic BMEC 'cobblestones' were exhibited under the microscope on the 10th day of culture. Meanwhile a relative lack of pinocytic vesicles and the tight junction between the BMEC were presented. BMEC displayed a significant restriction of paracellular transport. CONCLUSIONS: An in vitro BBB model by co-culture with allogenic BMEC with AS was established, which was simple, reliable and reproducible as well as similar to the in vivo BBB in respect to morphology, ultrastructures and restriction characteristics.
No abstract available
