Age Factors ; Animals ; Animals, Newborn ; Blood-Brain Barrier ; Brain ; blood supply ; physiopathology ; Brain Ischemia ; physiopathology ; therapy ; Models, Animal ; Rats ; Stem Cell Transplantation

Objective To investigate the clinical characteristics and diagnostics of Kozlowski type spondylometaphyseal dysplasia (SMDK). Methods The clinical features, laboratory tests and genetic testing of one SMDK case were analyzed. Re-sults A eight-year-old male patient had more than 6 years course of disease. The clinical manifestations were stubby limbs, ifn-gers and toes, varus deformity, knee valgus deformity, scoliosis and lordosis and severe metaphyseal changes. The heterozygous mutations were detected in TRPV4 and NXX3-2 genes. Conclusions Typical clinical features combined with genetic diagnosis facilitate early detection and accurate diagnosis of SMDK.

Hematopoietic stem cell transplantation(HSCT) is one of the measures therapy on acute leukemia in children.Very-high-risk children with acute lymphocytic leukemia (ALL),such as Philadelphia chromosome-positive(Ph+),T cell immunophenotype,achieved the first complete remission(CR1),and then receive allogeneic HSCT(allo-HSCT).The transplantation group had significantly improved disease-free survival(DFS)and overall survival(OS) compared with chemotherapy group. ALL children in the second complete remission(CR2) were recommended to receive allo-HSCT,especially early-relapse group. All children in the third complete remission (CR3) were recommended to receive an allo-HSCT,and reduce replapse rate through inducing graft versus leukemia.Total body irradiation-based conditioning regimens in children with ALL has advantage over chemotherapy conditioning regimens.Children with acute myeloblastic leukemia (AML) achieved CR1 and then undergo all-HSCT.This group had significantly improved DFS and OS compared with chemotherapy group.The children with relapsed AML,if having suitable donor,were recommended to receive allo-HSCT. The children with AML who underwent transplantation relapse,and can receive the second transplantation.There was no advantage in children patients with AML using TBI-based conditioning regimens.Conditioning regimens consisted of high-dose cytarabine,in combination with granulocyte colony-stimulating factor has advantage,and improve DFS. Unrelated umbilical cord blood has emerged as a potential option.Compared with unrelated bone marrow transplantation,there was no different in DFS and relapse.For the extent,it is one of the donor hematopoietic stem cell sources.

The levels of plasma immunoreactive atrial natriuretic polypeptide (ir-ANP) were determined by radioimmunoassay in 59 children with congenital heart disease (CHD). It was showed that the ir-ANP levels of the patients with left to right shunt were higher than those of control (P0.05). The iir-ANP levels of patients with HYHA II degree were increased significantly (P

BACKGROUND: Cell labeling and nuclear magnetic resonance can non-invasively in vivo label the region, existing mode and some bionomics of transplanted neural stem cells (NSCs). OBJECTIVE: To observe the outcome of superparamagnetic iron oxide in vitro labeled human NSCs. DESIGN, TIME AND SETTING: The cytology, in vitro, observation study was performed at the Laboratory of Department of Pediatrics, Navy General Hospital of Chinese PLA from December 2006 to June 2007. MATERIALS: Aborted human embryo was provided by Navy General Hospital of Chinese PLA. Superparamagnetic iron oxide (Lot number 97060601) was produced by Advanced magnetics,inc., USA. METHODS: Monoplast suspension was isolated from human embryo hippocampus using the mechanical method, and in vitro incubated in NSC medium, supplemented with epidermal growth factor and basic fibroblast growth factor. 11.2 g/L superparamagnetic iron oxide was diluted into 28 mg/L using NSC medium, mixed with 1.0 mg/L polylysine (8 ?L), and then made into superparamagnetic iron oxide-polylysine composite labeled medium. NSC spheres with active proliferation were obtained, made into single cell suspension (1?109/L), and then treated with serum-free medium containing superparamagnetic iron oxide. One week later, various cytokines were removed, and 5% fetal bovine serum was used for 24 hours to induce the NSC differentiation. MAIN OUTCOME MEASURES: Prussian blue staining was utilized to determine marking positive rate. Immunofluorescence staining was applied to detect glial fibrillary acidic protein and neurofilament expression. RESULTS: Superparamagnetic iron oxide labeled NSCs were yellow, and the speed of clone formation was not stepped down compared with the non-labeled cells. 20 hours following superparamagnetic iron oxide labeling, blue iron particles in NSCs cytoplasm were found, with the positive rate of 90%. Following induction, superparamagnetic iron oxide labeled NSCs were positive for glial fibrillary acidic protein and neurofilament. CONCLUSION: Superparamagnetic iron oxide can highly effectively label NSCs in vitro, and not affect biological features following labeling. NSCs can normally amplify and orientedly differentiate into neurons and astrocytes.

To observe the changes of artrial natriuretic peptide (ANP) and endothelin (ET) lev-els in patients with c0ngenital heart disease (CHD) and discuss their mechanism and clinical significance.Methods: The pulmonary pressure of the pulmonary hypertension (PH)group was higher than 2 kPa- Thepulmonary pressure of Non-PH group was n0rmal. Healthy children served as control group. Results: (1)Plasma ANP level of the PH group was apparently higher than that 0f the N0n-PH group and c0ntrolgroup. (2) Plasma ET levels in the peripheral vein, vena cava,right atrium, right ventricle, pulmonaryartery and left atrium of the PH group were particularly higher than that of the Non-PH group and c0ntr0lgroup, especially in the left atrium. C0nclusi0n: (l) CHD with PH induces the increase in cardiac v0lumeand pressure load, thus accelerates the release 0f ANP. It is actually a compensatory mechanism for the el-evation of endogenic ANP. (2) ET is a vasoconstrictor. ET owns the importance at PH pathogenesis andhas a certain clinical value in the diagn0sis 0f PH. (3) ET causes ANP releasing, then inhibits ET effectthrough negative feed. B0th of them are antag0nists each other.

Objective To isolate and culture the neural stem cells (NSCs) from the cerebral cortex of newborn rats, and investigate the cell-replace responses of the NSCs transplanted into the sibling rats with focal cerebral cortex ischemic lesion. Methods The serum-free medium DMEM/F12 (1∶1) containing basic fibroblast growth factor (bFGF or FGF2) and epidermal growth factor (EGF) was used to culture the neural stem cell spheres. The NSCs were identified by detecting the neural stem cell marker nestin with enzyme immune assay and inducing neural stem cell spheres to differentiation. The NSCs which would be used in the transplantation experiment were labeled by BrdU incorporation when cultured in vitro. The focal ischemic models were made by opening the skulls and removing the cerebral menings of 4-day-old rats to stop the blood supply for the neopallium. The BrdU-labeled NSCs were transplanted into the cerebral lesion boundary zones of the focal ischemic sibling rat models. The experiment rats were divided into lesion-transplantation group, lesion-control group and sham-operation-control group. Recipients were killed and the brains were examined by detecting the BrdU-labeled cells with enzyme immunohistochemistry at 4,7,14,30 days postgrafting, indicating the grafts living and migration in the host. Results The neural stem cell spheres, which floated and grew in medium, expressed nestin, as well as gave rise to neurons and astrocytes, could be obtained through culturing the cells derived from the cerebral cortex of newborn rats in vitro for a week. In the transplantation of the NSCs, the grafts were easy to migrate along the boundary zones of the focal ischemic lesions, and promoted the restore of the tissue structures in the damaged areas, the damage recovered well through the cell-replace responses. The BrdU-labeled positive cells in the lesion areas were full of the visual fields under microscope, the greatest density of the positive cells were focused in the granular layer of the injured cerebral cortex and not found in remote sites from the lesion. The number of BrdU-labeled cells gradually decreased in the brains of the sham-operation-control rats, only a few positive cells were found when examined at 14 days postgrafting, significantly less than that in the lesion-transplantation rats. Conclusions NSCs exist in the cerebral cortex of newborn rats. The ischemia can promote proliferation and graft of NSCs. The grafted NSCs play an important role in the recovery of focal cerebral cortex ischemic lesion.

BACKGROUND:Lingo-1 has been identified as a negative regulator of oligodendrocyte differentiation and myelination, which may be closely related to the white matter damage, but there is no systematic report on the dynamic changes of Lingo-1 after white matter damage.
OBJECTIVE:Toexplore the dynamic expression of Lingo-1 at different time points after white matter injury in newborn rats.
METHODS:Seventy-eight Sprague-Dawley rats aged 3 days old were equaly and randomly divided into sham operation group and model group. In the model group, models of white matter injury were established by unilateral ligation of the right common carotid artery combined with hypoxia. In the sham operation group, the right common carotid artery was isolated only, without ligation or hypoxia.
RESULTSAND CONCLUSION:At 7 days after model induction, hematoxylin-eosin staining and immunohistochemical staining for myelin basic protein showed that a selective white matter injury was seen at the injury site of a rat model, suggesting successful model establishment. Fluorescent quantitative PCR and western blot assay results demonstrated that the expression levels of Lingo-1 mRNA and protein were significantly up-regulated at 1 day and reached a peak at 7 days post-surgery. After 7 days, above expression wasgradualy decreased and the up-regulation of Lingo-1 protein lasted to the 28 days post-surgery compared to the sham operation group. These results show that Lingo-1 protein was closely related to the brain white matter injury.

Objective To explore the efficacy of unrelated cord blood transplantation treatment of mucopolysaccharidosis Ⅰ (MPS Ⅰ).Methods A 4-year-and-2-month-old boy with MPS Ⅰ who received treatment of human leucocyte antigen-mismatched unrelated cord blood stem cell transplantation after diagnosis was identified.The pre-treatment regimen was Busulfan + Cyclophosphamide + Fludarabine (Bu/Cy4 + Flud).Bu with the dosage of 1.2 mg/kg,once every 6 hours,4 days;Cy with the dosage of 50 mg/(kg · d) for 4 days and Flud with the dosage of 30 mg/(m2 · d) lasted for 4 days,respectively.The day that the graft was transplanted was defined as 0 day,days betore transplantation as negative days,days after transplantation as positive days.After pre-treatment,4.60 × 107/kg of cord blood nucleated cells and 3.05 × 105/kg CD34 positive cells were transplanted into the child.The combination of Antihuman thymocyte globulin,Cyclosporin A and Mycophenolate mofetil was administrated for prophylaxis of graft versus host disease(GVHD).After transplantation,the patient was given granulocyte colony stimulating factor to promote reconstitution of hematopoiesis.Results The myeloid and platelet engraftment time was respectively 15 days and 24 days after transplantation.Short tandem repeat (STR) DNA fingerprinting showed a full donor chimerism on day 21 after transplantation,and the full donor chimerism was stable afterwards.The peripheral-blood α-L-iduronidase (IDUA) activity returned to the normal value,and the IDUA gene sequencing did not demonstrate any mutation in 83 days after transplantation.On day 12 after transplantation,pulmonary infection with pulmonary hypertension occurred.Grade-Ⅱ acute intestinal GVHD occurred on day 15,Grade-Ⅱ acute cutaneous GVHD on day 51,and chronic GVHD (cutaneous,localized) on day 180.Otherwise,the patient complicated with hemorrhagic cystitis on day 35.These complications was cured favourably.In an 18-month-follow-up,the height of the boy increased by 3 cm,and his body weight had increased by 2.4 kg.His corneas regained clear,and his hepatosplenomegaly disappeared.The glycosaminoglycan of urine was negative.The neurocognitive performance of the boy had a little improvement.The abnormalities of fingers and other skeletons had no marked change.Conclusions Unrelated cord blood transplantation for MPS Ⅰ have definited effect.It is the first case report in China on treatment of MPS Ⅰ by unrelated cord blood transplantation.The researchers have accumulated some preliminary experience for future treatment of MPS Ⅰ by unrelated cord blood transplantation.