The correlation between the severity of hypoxic-ischemic brain damage in neonatal rats and subsequent abnormal neurobehavior / 中国新生儿科杂志

Objective:To assess the severity of hypoxic-ischemic brain damage (HIBD) in neonatal rats and predict the occurrence of subsequent neurobehavioral abnormalities after brain injury by scoring and magnetic resonance imaging (MRI).Methods:7-day-old of 60 Sprague-Dawley (SD) rats were randomly divided into control group (14 rats), sham operation group (14 rats) and HIBD model group (32 rats). HIBD model was established by right common carotid artery dissection with Rice-Vannucci method and hypoxia. Within 24 h after modeling, the rats in the model group were evaluated by general condition score and Longa score, and the surviving rats with moderate and severe HIBD were selected for the experiment. 24 h after modeling, 5 rats of the model group were randomly selected for 2,3,5-triphenyltetrazole chloride staining to verify cerebral infarction. 1 week after modeling, 6 rats from each group were randomly selected for hematoxylin-eosin staining to observe HIBD brain injury. 4 weeks after modeling, 4 rats were randomly selected from the control group and the sham operation group, and 8 rats from the remaining model group were used to evaluate the volume of brain damage by MRI. 5-6 weeks after modeling, the remaining 8 rats from each group were subjected to the Cylinder test, and at 13 weeks, they underwent the Morris water maze test to evaluate their neurobehavior.Results:In HIBD model group, 19 rats with moderate to severe HIBD were selected from 32 rats. 24 h after modeling, cerebral infarction was verified in all rats, indicating moderate to severe HIBD. Brain tissue pathology observed 1 week after modeling revealed predominantly gray matter brain damage. MRI showed that 7 out of 8 rats had moderate to severe HIBD. Compared to the control and sham operation groups, the model group exhibited a significant decrease in the usage rate of the left forelimb in the Cylinder test at 5-6 weeks after modeling ( P<0.05), and the latency period in Morris water maze test was significantly prolonged at 13 weeks after modeling ( P<0.05), and the times of crossing platform quadrant were significantly reduced ( P<0.05). There was no significant difference in the right brain injury volume between 24 h and 4 weeks model group ( P>0.05). The brain injury volume in model group was negatively correlated with the usage rate of left forelimb in cylinder test at 5-6 weeks and the times of crossing platform quadrant in Morris water maze test at 13 weeks ( P<0.05), and positively correlated with latency period in Morris water maze test at 13 weeks ( P<0.05). Conclusions:Within 24 h of HIBD modeling, the severity of brain injury can be preliminarily predicted by general condition score and Longa score. 4 weeks after modeling, in the chronic phase of brain injury, MRI was proved to be an excellent predictor for mid-term and long-term neurobehavioral abnormalities in HIBD rats.

Protective effect of transplantation of human oligodendrocyte precursor cells in a rat model of white matter injury / 中国当代儿科杂志

OBJECTIVE@#To study the effect of human oligodendrocyte precursor cell (hOPC) transplantation in the treatment of white matter injury (WMI).@*METHODS@#Neonatal rats were randomly divided into a sham-operation group, a model group, and a transplantation group (@*RESULTS@#The place navigation test using the Morris water maze showed that the model group had a significantly longer escape latency than the sham-operation group, and compared with the model group, the transplantation group had a significant reduction in escape latency (@*CONCLUSIONS@#Intrathecal hOPC transplantation may alleviate neurological injury and promote remyelination in a rat model of WMI.

A clinical study of haploid hematopoietic stem cells combined with third-party umbilical cord blood transplantation in the treatment of chronic granulomatous disease / 中国当代儿科杂志

OBJECTIVE@#To investigate the clinical efficacy of haploid hematopoietic stem cells (haplo-HSC) combined with third-party umbilical cord blood (tpCB) transplantation in the treatment of X-linked chronic granulomatous disease (X-CGD).@*METHODS@#The clinical data of 26 boys with X-CGD were retrospectively analyzed who were admitted to the Sixth Medical Center of PLA General Hospital between April 2014 and March 2018. All the patients were treated with haplo-HSC combined with tpCB transplantation. The median age of the patients was 3.5 years. The donor was the father in 25 cases and an aunt in 1 case. Transplantation was 5/6 HLA-matched in 9 cases, 4/6 in 12 cases, and 3/6 in 5 cases. The patients received busulfan, cyclophosphamide, fludarabine, or anti-thymocyte globulin for myeloablative preconditioning. Cyclosporine A and mycophenolate mofetil were used for prevention of acute graft-versus-host disease (aGVHD). Then the patients were treated with haploid bone marrow hematopoietic stem cells combined with tpCB transplantation on day 1 and haploid peripheral hematopoietic stem cells on day 2. The counts of median donor total nucleated cells, CD34 cells, and CD3 cells were 14.6×10/kg, 5.86×10/kg, and 2.13×10/kg respectively.@*RESULTS@#The median time to neutrophil and platelet engraftment was 12 and 23 days after transplantation respectively. Full donor hematopoietic chimerism was observed on day 30. Twenty-five cases were from haplo-HSC and 1 was from cord blood. No primary implant failure and implant dysfunction occurred, and secondary implant failure occurred in one case. The NADPH oxidase activity returned to normal one month after transplantation. The incidence of grade I-II aGVHD and grade III-IV aGVHD was 35% and 15% respectively. Chronic GVHD (cGVHD) of the skin occurred in one case, and no progression was observed after steroid administration. During the follow-up period of 6-51 months, 25 patients survived, of whom 24 were disease-free (23 patients without cGVHD and 1 with cGVHD of the skin) and NADPH oxidase activity returned to normal; one patient developed secondary implant failure but survived; one patient died of viral interstitial pneumonia 16 months after transplantation. The 5-year event-free survival rate and overall survival rate were 81%±12% and 89%±10% respectively.@*CONCLUSIONS@#Haplo-HSC combined with tpCB transplantation is one of the effective methods for the treatment of X-CGD in children.

A clinical study of haploidentical hematopoietic stem cell transplantation in the treatment of pediatric patients with acquired severe aplastic anemia: single center experience / 中华血液学杂志

Objective@#To investigate the efficacy of haplotype hematopoietic stem cell transplantation in the treatment of acquired severe aplastic anemia (SAA) in children.@*Methods@#The clinical characteristics of 59 pediatric patients with SAA, including 26 cases VSAA, 37males and 22 females, 47 cases typeⅠ and 12 cases typeⅡ, undrerwent haplo-HSCT in our hospital between December 1st, 2011 and December 1st, 2017 were retrospectively analyzed. Among 59 patients, 56 patients with a median age of 4.5 (1.2-14.8) years and median weight of 43 (12-80) kg underwent their first HSCT and 3 patients underwent their second HSCT. All patients received the following conditioning regimen: busulfan, cyclophosphamide, and rabbit ATG or Bu (–, CTX) , fludarabineand rabbit ATG. The prophylaxis of acute graft versus host disease (aGVHD) was cyclosporine (CsA) , MMF and methotrexate. All patients received bone marrow transfusion on day 01 and peripheral stem cell transfusion on day 02 from haploid donor. The median dose of donor mononuclear cell counts was 15.60 (7.74-21.04) ×108/kg of recipient weight and CD34+ cell counts was 4.86 (3.74-7.14) ×106/kg of recipient weight.@*Results@#Neutrophils and platelets of all 59 children were implanted. The median implantation time of granulocytes and platelets were 13 (10-19) d, 19 (9-62) d, respectively. The incidence of grade Ⅰ-Ⅱ aGVHD was 45.76% (27 cases) and grade Ⅲ/Ⅳ 13.56% (8 cases) , The incidence of chronic GVHD was 8.47% (5 cases) , The incidences of CMV and EBV viremia were 59.32% (35 cases) and 28.81% (17 cases) , respectively. The median follow-up was 30 (8-80) months, 57 patients survived with disease free, 2 patients died of GVHD. Both of the estimated 5-year OS and DFS rates were (96.4±2.5) %.@*Conclusion@#Haplo-HSCT could improve the outcomes of SAA children.

A comparative study on the secretion of various cytokines by human adipose-derived stem cells at different passages / 中国组织工程研究

BACKGROUND: Adipose-derived stem cells (ADSCs) can establish a favorable repair microenvironment by secreting abundant cytokines, which allows ADSCs to be a good source of seed cells for the treatment of ischemic diseases. OBJECTIVE: To investigate the changes of cytokines secreted by human ADSCs at passages 2-10. METHODS: After isolation and culture of ADSCs from human adipose tissue, the morphological features of cells were observed under inverted microscope. Human ADSCs were identified by the immunophenotypes and differentiation capability. RESULTS AND CONCLUSION: ADSCs were fusiform or polygonal in shape, with buging cell body, homogenized cytoplasm and clear nuclei, and could differentiate into adipocytes, osteocytes and chondroblasts in vitro. ADSCs at passage 3 were positive for CD29 (99.21%), CD73 (99.65%) and CD90 (99.92%), but negative for hematopoietic marker CD34 (2.25%). ELISA results showed that ADSCs at passage 5 had the highest secretion levels of vascular endothelial growth factor and hepatocyte growth factor, while ADSCs at passage 3 had the highest secretion level of brain-derived neurotrophic factor. To conclude, ADSCs have steady biological features of stem cells and exhibit good growth and proliferation potentials. ADSCs at different passages have different capacities in the secretion of vascular endothelial growth factor, hepatocyte growth factor and brain-derived neurotrophic factor. Passage 5 ADSCs show the highest ability to secrete vascular endothelial growth factor and hepatocyte growth factor, while passage 3 ADSCs show the strongest potential to secrete brain-derived neurotrophic factor.

Safety and early curative efficacy of lipocyte bioactive secretion in preventing white matter injury of premature infants / 中华实用儿科临床杂志

Objective To explore the safety and efficacy of intrathecal administration of adipose stem cells de-rived from bioactive secretome (ASCBS)in treatment of whiter matter injury (WMI)in the preterm infants. Methods Sixty - three cases of WMI were recruited according to the uniform standards from multiple medical centers and they were divided into 3 gestational age (GA)subgroups,which were 21 cases in group A (GA 24 - 28 + 6 ),20 cases in group B (GA 29 - 32 + 6 ),and 22 cases in group C (GA 33 - 36 + 6 ). The patients were randomly divided into treatment groups and control groups by tossing coins. The treatment groups received lumbar puncture followed with ASCBS intra-thecal injection once daily for 3 consecutive days. Follow - up study included Neonatal Behavioral Neurological Assess-ment (NBNA)at term - equivalent age and neurodevelopment at corrected age of 6 - month. Neurodevelopment was assessed by using the Bayley Scales of Infant Development and Peabody Developmental Motor Scale. The survival rates, NBNA scores,mental development index (MDI),psychomotor develop index (PDI),total motor development quotient, gross motor development quotient and fine motor development among each subgroup were compared. Results Sixty -three cases were recruited,including 31 in the treatment group and 32 in the control group. Only 1 case in the treatment groups lost in the follow - up. No clinical side effects were found in the treatment groups. There was no significant diffe-rence in the survival rate and complication in the preterms in all subgroups of the treatment group and control group (all P > 0. 05). The gross and total motor development quotient in the treatment group A was higher than that in the control group A(gross motor development quotient:98. 330 ± 6. 282 in treatment group A,90. 330 ± 3. 777 in control group A, P = 0. 040;total motor development quotient:97. 330 ± 4. 803 in treatment group A,91. 000 ± 4. 472 in control group A,P = 0. 023). The rest findings showed no significant difference between groups. Conclusion The treatment of WMI in preterm infants with ASCBS is safe and can promote the motor development of preterm infants with GA in 24 - 28 weeks.

Curative effect of human umbilical cord mesenchymal stem cells treatment on refractory acute graft versus host disease of children after allogeneic hematopoietic stem cell transplantation / 中华实用儿科临床杂志

Objective To investigate the curative effect and safety of human umbilical cord mesenchymal stem cells(UCMSCs)on the treatment of refractory acute graft versus host disease(aGVHD)of children after allogeneic hematopoietic stem cell transplantation.Methods Five children with refructory aGVHD who hospitalized at the De-partment of Pediatrics,the Navy General Hospital were treated with UCMSCs retrospectively.Among them,1 case was male and 4 cases were female,who aged from 18 months old to 15 years old.Two cases had aplastic anemia(AA),1 case with acute myeloblastic leukemia(AML-M2-CR2),1 case with acute lymphoblastic leukemia(ALL-CR1)and 1 case with myelodysplastic syndrome(MDS). Three cases received peripheral blood stem cell transplantation from HLA-matched sibling donor,1 case received mother′s peripheral blood and bone marrow stem cell transplantation from the haploid donor,and 1 case received father′s peripheral blood and bone marrow stem cell transplantation from the haploid donor(both 3/6 HLA locus matched).Prophylaxis for graft versus host disease(GVHD)was performed by using ciclosporin A and methotrexate in 1 case,others used anti-thymocyte globulin,ciclosporin A,mycophenolate mofetil and methotrexate for GVHD prophylaxis. All children developed refractory aGVHD,and they received 3-5 kinds of immunosuppressive agents to treat the refractory aGVHD,but the therapeutic effect was very poor.Then the children received UCMSCs infusion 2 or 3 times(once a week),the UCMSCs dose given was(1.5-2.0)×106per kg body weight,the curative effect and adverse reactions were apparent after infusion.Results All the children with re-fractory aGVHD were improved after treatment,the overall response was 100%,and 2 cases were healed and dis-charged,1 case suffered from relapsed of aGVHD and died,and the other 2 cases suffered from relapsed of aGVHD and died of thrombotic microvascular disease.On adverse reaction was monitored during infusion,and 2 cases had disease-free survival during 2 years follow-up,without tumour and primary disease recurrence.Conclusions UCMSCs is safe and effective for treatment of refractory aGVHD.In order to improve the curative effect and disease-free survival,the UCMSCs should be reduced early,which can reduce the application and side effects of the immunosuppressor.

A comparative study on autologous and allogeneic adipose derived mesenchymal stem cell transplantation in treatment of acute myocardial infarction rats / 解放军医学杂志

Objective To investigate the effects of autologous and allogeneic adipose derived mesenchymal stem cells (ADMSCs) transplantation on rat model of acute myocardial infarction (AMI) and possible mechanisms.Methods The AMI models were established with 45 male Lewis rats by ligation of left anterior descending coronary artery,and then randomly divided into 3 groups (15 each) including AMI group,allogeneic ADMSC transplantation group (Allo-ADMSC group) and autologous ADMSC transplantation group (Auto-ADMSC group).After successfully modeling,CM-Dil-labeled third-generation ADMSCs (2 × 106) were implanted into the myocardium of rats within 1 hour,and rats in AMI group were injected with equal amount of PBS.The cardiac function,immunofluorescence and Masson were identified 4 weeks after transplantation.Results Four weeks after transplantation,compared with AMI group,the left ventricular ejection fraction in Allo-ADMSC group and Auto-ADMSC group increased significantly,the left ventricular end-systolic and end-diastolic diameter decreased,the collagen deposition fraction decreased significantly (P＜0.05).Compared with Allo-ADMSC group,the left ventricular ejection fraction increased in AutoADMSC group,the number of newborn capillaries increased and the myocardial collagen deposition fraction decreased (P＜0.05).Conclusion Autologous ADMSC can promote vascular proliferation in the infarct area more better than allogeneic ADMSC,reduce the local collagen deposition,extenuate the degree of myocardial fibrosis,thereby to inhibit collagen remodeling,and repair damaged myocardial tissue and improve heart function.

Protective effect of miconazole on white matter damage induced by anoxia and ischemia in rats / 临床儿科杂志

Objective To explore the protective effect of miconazole on white matter damage (WMD) in neonatal rats. Methods Three-day-old neonatal SD rats were randomly divided into sham group, WMD model group, 10 mg/(kg·d) miconazole group and 40 mg/(kg·d) miconazole group with 15 rats each. Rats in WMD model group were subjected to the ligation of right carotid artery, and then kept in a chamber with 6% oxygen and 94% nitrogen for 80 min to establish the white matter damage model. The rats in miconazole group were intraperitoneally injected with different doses (10 and 40mg/kg) of miconazole, dissolved in dimethyl sulfoxide (DMSO), for five consecutive days, and rats in WMD model group were injected with the same volume of DMSO. Myelin basic protein (MBP) of white matter was detected by immunofluorescence staining and western blot. Myelin sheaths of corpus callosum were observed by transmission electron microscopy. Weight changes of rats were compared among groups. Results Immunofluorescence staining and western blot showed that, after treatment with miconazole, the MBP expression level of corpus callosum was higher than in WMD model group (P<0.05). In WMD model group, the myelin sheath of corpus callosum had loose structure and a large number of small vacuoles with decreased thickness of myelin sheath. After treatment with miconazole, myelinolysis induced by anoxia and ischemia could be improved significantly. The increase in weight of rats in WMD model group was significantly less than that in sham group. And after miconazole treatment, the rate of weight gain of rats was increased. Conclusion Miconazole can significantly reduce the brain white matter damage induced by anoxia and ischemia through promoting myelination, and then improves the growth and development in rats.

Neuroprotective effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats / 中国新生儿科杂志

Objective To explore the effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats and to compare the difference of therapeutive effect between the cerebellum medulla oblongata pool injection and the jugular vein injection.Method A total of 73 three-day-old SD rats were chosen to establish the model of white mater injury.After 24 hours,the 73 rats were randomized into the experimental group (n =46) and the control group (n =27).Then the experimental group was reclassified into ventricular group (n =23) and intravenous group (n =23).In the ventricular group,the paracrine extracts of human adipose stem cells was injected locally into the cerebellum medulla oblongata pool injection,while the extracts was injected into the jugular vein in the intravenous group.The control group was reclassified ventricular control group (n =15) and intravenous control group (n =12),and equivoluminal saline was injected the same way as the experimental group.Frozen sections of the brain tissue from 3 rats of each experimental group one day after injection were stained with fluorescein-conjugated streptavidin to study the distribution of the extracts.The brain tissue of 3 rats from each subgroup 3 days after injection were stained with hematoxylin eosin (HE) to observe the pathomorphological changes.While 7 days later,myelin basic protein (MBP) of white matter which was obtained from 7 rats of each group was detected by immunofluorescence staining.28 days after injection,the remaining rats were assessed by neurobehavior tests.For the rats that died during the experiment,the same number of the rats would be substituted in this study.Result The paracrine extracts were found to transfer to the brain lesion area,and the amount of the extracts was more in the ventricular group.The results of the HE staining showed that the white matter injury was more severe in the ventricular control group,and extensive area of infarction were found in this group.White matter injury was mild in the experimental group,and the structure of the corpus callosum was more complete in the ventricular group.MBP semi-quantitative scores of the ventricular group (0.7 ± 0.3) and intravenous group (1.7 ± 0.3) were lower than those of ventricular control group (3.4 ± 0.4)and intravenous control group(3.3 ±0.3).And the MBP scores of ventricular group was significantly lower than that of intravenous group (P ＜ 0.05).The scores of the neurobehavioral tests of the experimental group were significantly higher than those of the control group,while the scores of the ventricular group were significantly higher than those of the intravenous group (P ＜ 0.05).Conclusion The paracrine extracts derived from adipose stem cells could improve the prognosis of white matter injury,and cerebellum medulla oblongata pool injection showed better curative effect than the jugular vein injection.

Culture and identification of endothelial progenitor cells from human adipose tissue with CD31 immunomagnetic beads / 中国组织工程研究

BACKGROUND:Endothelial progenitor cells are precursor cells of mature endothelial cells,which can migrate to ischemic tissues and differentiate into mature endothelial cells,and then play an important role in vascular remodeling.Endothelial progenitor cells have wide application prospects in various ischemic diseases,but the biological characteristics and identification methods are still controversial.OBJECTIVE:To investigate the methods of isolation and culture of endothelial progenitor cells from the human adipose tissue and to identify their biological features,in order to provide a sufficient source of cells for ischemic diseases.METHODS:Stromal vascular fraction cells were isolated from the human adipose tissue by enzymatic digestion,CD31+ cells were selected using immunomagnetic beads,and then cultured in endothelial basal medium-2 supplemented with the EGM-2-MV-SingleQuots.Endothelial progenitor cells were identified through detection of morphology,cell markers and cell functions.RESULTS AND CONCLUSION:(1) CD31 + cells were selected by immunomagnetic beads and then cultured and amplified in vitro,which displayed typical cobblestone-like morphology,and they maintain their proliferative ability.(2) Flow cytometry results showed that the CD31+ cells expressed CD31 (98.84％),CD34 (97.21％),VEGRR2 (64.07％),CD146 (98.42％) and CD133 (2.55％),but hardly expressed CD45 (1.1％),a hematopoietic stem cell marker.(3) The CD31 + cells were also found to incept Dil-ac-LDL and exhibit lectin binding capability.Furthermore,a lumen-like structure was formed in Matrigel,which has the ability of angiogenesis in vitro.To conclude,these results suggest that it is feasible to isolate and culture endothelial progenitor cells from the human adipose tissue by enzymatic digestion combined with immunomagnetic bead sorting.

Long-term effect of oligodendrocyte precursor cell transplantation on a rat model of white matter injury in the preterm infant / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the long-term effect of oligodendrocyte precursor cell (OPC) transplantation on a rat model of white matter injury (WMI) in the preterm infant.</p><p><b>METHODS</b>A total of 80 Sprague-Dawley rats aged 3 days were randomly divided into sham-operation group, model control group, 5-day ventricular/white matter transplantation group, 9-day ventricular/white matter transplantation group, 14-day ventricular/white matter transplantation group (n=10 each). All groups except the sham-operation group were treated with right common carotid artery ligation and hypoxia for 80 minutes to establish a rat model of WMI in the preterm infant. OPCs were prepared from the human fetal brain tissue (10-12 gestational weeks). At 5, 9, and 14 days after modeling, 3×10OPCs were injected into the right lateral ventricle or white matter in each transplantation group, and myelin sheath and neurological function were evaluated under an electron microscope at ages of 60 and 90 days.</p><p><b>RESULTS</b>Electron microscopy showed that at an age of 60 days, each transplantation group had a slight improvement in myelin sheath injury compared with the model control group; at an age of 90 days, each transplantation group had significantly thickened myelin sheath and reduced structural damage compared with the model control group, and the 14-day transplantation groups had the most significant changes. There were no significant differences in the degree of myelin sheath injury between the ventricular and white matter transplantation groups at different time points. At an age of 60 or 90 days, the transplantation groups had a significantly higher modified neurological severity score (mNSS) than the sham-operation group and a significantly lower mNSS than the model control group (P<0.05).</p><p><b>CONCLUSIONS</b>OPC transplantation may have a long-term effect in the treatment of WMI in the preterm infant, and delayed transplantation may enhance its therapeutic effect.</p>

Dynamic expression of Lingo-1 in a model of white matter injury / 中国组织工程研究

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.

Treatment of Gaucher disease with allogeneic hematopoietic stem cell transplantation: report of three cases and review of literatures / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore the efficacy of unrelated umbilical cord blood transplantation (UCBT) in the treatment of Gaucher disease.</p><p><b>METHOD</b>The clinical characteristics of three children with Gaucher disease underwent UCBT in our hospital between April 2013 and September 2014 were retrospectively analyzed. Literature on allogeneic hematopoietic stem cell transplantation (allo-HSCT) in the treatment of Gaucher disease was searched at Wanfang and Pubmed databases between 1983 and 2015 and was reviewed and summaried.</p><p><b>RESULT</b>Three children with Gaucher disease, all were female, received UCBT. These patients' age at receiving transplantation was 3.8 years, 7.1 years and 2.6 years, respectively. The second case received the second transplantation. The first and third case received splenectomy before UCBT. The pretreatment regimen was busulfan (Bu)/fludarabine (Flu)/cyclophosphamide (CTX)/antithymocyte globulin (ATG), and for the patient received the second transplantation melphalan was added to the myeloablative conditioning regimen of Bu/Flu/CTX/ATG. Cyclosporine and mycophenolate mofetil (MMF) wee used for prophylaxis of acute graft versus host disease (aGVHD). The dose of cord blood stem cell nucleated cell counts was 9.7 × 10⁷ /kg,11.9 × 10⁷ /kg and 7.6 × 10⁷/kg respectively. The dose of cord blood stem cell CD34⁺ cell counts was 5.4 × 10⁵/kg , 3.5 × 10⁵/kg and 3.2 × 10⁵/kg respectively. The day of granulocytes exceeding 0.5 × 10⁹/L was day 11, 12 and 19 after transplantation, respectively. The day of platelets exceeding 20 × 10⁹/L was day 14, 33 and 74 after transplantation, respectively. At one month after transplantation the rate of chimerism was over 95% and all patients got donor complete chimerism. The level of β-glucocerebrosidase recovered to normal at one month after transplantation. During transplantation, all patients developed cytomegalovirus (CMV) and Epstein-Barr virus (EBV) viremia. In case 1 immune thrombocytopenia occurred at five month after transplantation unresponding to steroids and mesenchymal stem cells infusion was administered and his platelet in routine blood test recovered to normal. But the patient died because she was infected with varicella-zoster virus out of hospital at nine month after transplantation and the level of β-glucocerebrosidase was normal before death and chronic GVHD (cGVHD) was not found. The case 2 is now in 19th month after transplantation and his level of β-glucocerebrosidase was normal. cGVHD was not found. The patient is currently free of disease. The case 3 was in 9th month after transplantation and his level of β-glucocerebrosidase was normal. cGVHD was found at 112 day after transplantation and was localized and could be controlled by hormonal therapy. The patient is currently free of disease. Three patients' size of liver was significantly reduced after their level of β-glucocerebrosidase ecovered. There were 50 cases with Gaucher disease who were treated with allo-HSCT in the literature and none of them were reported from China. Disease-free survival rate of patients treated with allo-HSCT for Gaucher disease was 85%. In all reports, there were 31 cases who had information of typing of Gaucher disease, of whom 22 cases had type 1 and 9 cases had type 3. Twenty-nine cases had information of survival, of whom 24 cases survived and 5 cases died of infection. Fifteen cases had data of engraftment, 2 of whom had graft failure and one had late graft failure.Glucocerebrosidase recovered to normal in 25 of 31 cases who had relevant data, in one of whom with late graft failure the enzyme recovered to normal 3 month after transplantation, but his enzyme decreased to the initial level 9 month after transplantation. Along with enzyme level's recovery to normal, in a part of cases bone pain and hepatomegaly were relieved and growth delay was improved.</p><p><b>CONCLUSION</b>The unrelated UCBT may be a form of treatment that offers the potential of permanent cure and a procedure with possible long-term benefits in patients with Gaucher disease.</p>

Kozlowski type spondylometaphyseal dysplasia:one case report / 临床儿科杂志

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.

Forty cases of cerebral palsy treated with human neural stem cell transplantation / 中华行为医学与脑科学杂志

Objective To assess the clinical efficacy of human neural stem cell (hNSC) transplantation in the treatment of severe cerebral palsy (CP) in children.Methods hNSCs were obtained from the forebrain of 10 to 12-week-fetus.Forty children with CP were voluntarily received hNSC transplantation that were injected into cerebral ventricle.The development of motor and fine motor functions were evaluated by GMFM and PDMS-FM 1 month before hNSC transplantation.as well as 3 and 6 months after hNSC transplantation.Results Twenty six (65％) cases displayed improvement from day 5 to month 6 after hNSC transplantation.GMFM assessment showed that the percentage was (4.52±2.50) ％ 1 month before hNSC transplantation,(7.74±2.94) ％ 3 months after hNSC transplantation and (13.01±6.71)％ 6 months after hNSC transplantation,indicating a significant improvement by the treatment of hNSC transplantation(P＜0.05).The percentage in PDMS-FM evaluation was (15.01± 12.00)％,(20.34± 11.91) ％ and (30.02± 12.50) ％ one month before hNSC transplantation,3 and 6 months after hNSC transplantation,respectively,also suggesting a significant improvement induced by hNSC transplantation treatment (P＜0.05).Moreover,the developmental improvement was the most prominent among 1-3 months post hNSC transplantation.Then the development slowed down.Significantly,patients received no hNSC transplantation experienced serious adverse events or complications.Conclusions hNSC transplantation is an effective and safer therapy for severe CP.Future observations are needed to evaluate long-term clinical efficacy of the therapy.

The effect of different hypoxic duration on the degree of brain white matter injury / 临床儿科杂志

ObjectiveTo explore the effect of different hypoxic duration on the brain white matter injury.Methods ewborn rats were randomly divided into two groups, normal group (n=24) and model group (n=45). The model group was di-vided into 3 subgroups (n=15) according to the time of hypoxia (50 min, 70 min, and 90 min). The animal model of white matter injury was established by unilateral carotid artery ligation in model group. After different duration of hypoxia, the mortality rate was recorded, the morphological changes of brain pathology was observed by hematoxylin eosin (HE) staining, myelin basic pro-tein (MPB) of white matter was detected by immunolfuorescence staining and motor function was evaluated by climbing slope test.ResultsThe mortality rates signiifcantly increased with prolonged hypoxia. The mortality rate was as high as 60% in 90 min subgroup. The HE staining showed that there were no obvious injury in 50 min subgroup, selective white matter injury on the operative side appeared in 70 min subgroup, and a wide range of infarction of white matter, hippocampus, and cortex appeared in 90 min subgroup. MBP semi-quantitative scores of white matter injury were higher in 70 min subgroup (3.89 ± 0.47) and 90 min subgroup (4.72 ± 0.57) than that in the normal group (0.06 ± 0.24), the difference was statistically signiifcant (P <0.05). In climb-ing slope test, the subgroups had different degrees of motor dysfunction on affected side with 90 min subgroup being the most serious.ConclusionsWhite matter injury model could be established by unilateral carotid artery ligation, and different hypoxic duration signiifcantly affects the range and degree of injury.

Clinical research on unrelated cord blood transplantation treatment for mucopolysaccharidosis Ⅰ / 中华实用儿科临床杂志

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.

A long-term and economical method for culturing human oligodendrocyte precursor cells / 中华实用儿科临床杂志

Objective Cell therapy is a possible effective way to treat myelination disorder diseases.Cell therapy needs to apply a method for culturing oligodendrocyte precursor cells.Therefore,this study was to develop a stable,efficient and economical method for obtaining human oligodendrocyte precursor cells (OPCs) in order to provide cell required for clinical treatment.And this will provide a new option for clinical applications.Methods Human OPCs were obtained through magnetic bead sorting and cultured in OPCs proliferation medium.For a short-time (2,4,6,8days) and long-time culture,morphology of OPCs was observed.The fourth generation of OPCs was analyzed for expression of OPCs specific markers O4,Soxl0 and platelet derived growth factor alpla receptors(PDGFαR) and the capacity to differentiate into oligodendrocytes by immunofluorescence staining.At the same time,the effects of B27 and N1 on OPCs growth state were inspected as well.Results For a short-time culture,OPCs had typical bipolar or tripolar morphology and proliferated in good condition.For a long-time culture,all 4 generations OPCs had typical bipolar or tripolar morphology;the fourth generation OPCs highly(＞ 90％) expressed 04,Sox10 and PDGFαR,after induction,OPCs could be differentiated into oligodendrocytes.After 4 generations of long-time culture,OPCs already maintained the original sharp,high purity and had the capacity to differentiate into oligodendrocytes.It was indicated that this culture system was suitable for human OPCs for a long-time culture.Conclusions Overall,using this culture system,isolated human OPCs not only can be stably cultured and proliferated in vitro,but also have the capacity to differentiate into oligodendrocytes.From this reproducible method,a large number of human OPCs can be stably obtained in vitro as convenient as possible.And this will provide a new option for clinical applications.This method uses fewer cytokines.Therefore,this method will provide stable,efficient and economical OPCs for cell therapy of myelination disorders or myelin damage diseases.

Clinical effect of umbilical cord blood transplantation in 37 pediatric patients with hematologic malignancies: a single-center experience / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the clinical effect of umbilical cord blood transplantation (UCBT) in children with hematologic malignancies.</p><p><b>METHODS</b>A retrospective analysis was performed on the clinical data of 37 pediatric patients with hematologic malignancies that consisted of 14 cases of acute lymphocyte leukemia, 9 cases of acute myeloid leukemia, 5 cases of juvenile myelomonocytic leukemia, 3 cases of chronic myeloid leukemia, 2 cases of acute mixed leukemia, 3 cases of myelodysplastic syndrome, and 1 case of lymphosarcomatous leukemia. Thirty-seven children with hematologic malignancies received UCBT from unrelated donors (34 cases) and related donors (3 cases). Grafts were 6/6 HLA-matched in 5 cases, 5/6 HLA-matched in 12 cases, 4/6 HLA-matched in 11 cases, and 3/6 HLA-matched in 9 cases. Before transplantation, these patients received rabbit antithymocyte globulin-containing conditioning regimen. The myeloablative conditioning regimen was given in 36 cases and the reduced-intensity conditioning regimen in one case. The median age of transplantation was 5.7 years, and the median weight was 20 kg. The grafts that contained a median of 6.2×10(7) total nucleated cells (TNC)/kg and 2.7×10(5) CD34(+) cells/kg were infused.</p><p><b>RESULTS</b>The median times to neutrophil engraftment and platelet engraftment were 12 days and 25 days, respectively, and the rates of neutrophil engraftment and platelet engraftment were 95% and 78%, respectively. The rate of neutrophil engraftment was positively correlated with the number of CD34(+) cells (P=0.011), while the rate of platelet engraftment was correlated with the numbers of CD34(+) cells and TNC (P=0.001; P=0.014). The incidence rates of acute and chronic graft-versus-host disease were 49% and 11%, respectively. The median follow-up was 54 months. The 5-year transplant-related mortality, overall survival, and disease-free survival were 27%, 57.4% and 41%, respectively.</p><p><b>CONCLUSIONS</b>UCBT is an alternative source of hematopoietic stem cells for patients with hematologic malignancies.</p>