OBJECTIVE: To explore the effect of hypoxia-supported umbilical cord mesenchymal stem cell (UC-MSC) on the expansion of cord blood mononuclear cell （MNC） in vitro. METHODS: The isolated cord blood mononuclear cells were inoculated on the preestablished umbilical cord mesenchymal stem cell layer and cultured under hypoxic conditions (3％ O₂) and the experimental groups were normoxia (MNCs were cultured under normoxic conditions), hypoxia (MNCs were cultured under hypoxic conditions), UC-MSC (MNCs were cultured with UC-MSC under normoxic conditions), and UC-MSC+hypoxia (MNCs were cultured with UC-MSC under hypoxic conditions). To further investigate the combinational effect of 3 factors of SCF+FL+TPO (SFT) on expansion of cord blood MNCs in vitro in hypoxia-supported UC-MSC culture system, the experiments were further divided into group A (MNCs were cultured with UC-MSC and SFT under normoxic conditions), group B (MNCs were cultured with UC-MSC under hypoxic conditions), group C (MNCs were cultured with UC-MSC and SFT under hypoxic conditions). The number of nucleated cells (TNC), CD34⁺ cell, CFU and CD34⁺CXCR4⁺, CD34⁺CD49d⁺, CD34⁺CD62L⁺ cells of each groups were detected at 0, 7, 10 and 14 days, respectively. RESULTS: Compared with group hypoxia and UC-MSC, group UC-MSC+hypoxia effectively promoted the expansion of TNC, CD34⁺ cell and CFU, and upregulated the expression level of adhesion molecule and CxCR4 of the cord blood CD34⁺ cell（P<0.05）. After culturing for 14 days, compared with group A and group B, group C effectively promoted the expansion of cord blood MNC at different time points（P<0.05）, and the effect of group A was better than that of group B at 7 and 10 days（P<0.05）. CONCLUSION Hypoxia-supported UC-MSC efficiently promoted the expansion and expression of adhesion molecule and CXCR4 of cord blood CD34⁺ cell, and the effect of expansion could be enhanced when SFT 3 factors were added.
Humans ; Cells, Cultured ; Fetal Blood ; Cell Proliferation ; Umbilical Cord/metabolism* ; Mesenchymal Stem Cells ; Antigens, CD34/metabolism* ; Hypoxia/metabolism*

Preeclampsia and intrauterine growth restriction (IUGR) of the fetus are the two most common pregnancy complications worldwide, affecting 5%-10% of pregnant women. Preeclampsia is associated with significantly increased maternal and fetal morbidity and mortality. Hypoxia-induced uteroplacental dysfunction is now recognized as a key pathological factor in preeclampsia and IUGR. Reduced oxygen supply (hypoxia) disrupts mitochondrial and endoplasmic reticulum (ER) function. Hypoxia has been shown to alter mitochondrial reactive oxygen species (ROS) homeostasis and induce ER stress. Hypoxia during pregnancy is associated with excessive production of ROS in the placenta, leading to oxidative stress. Oxidative stress occurs in a number of human diseases, including high blood pressure during pregnancy. Studies have shown that uterine placental tissue/cells in preeclampsia and IUGR show high levels of oxidative stress, which plays an important role in the pathogenesis of both the complications. This review summarizes the role of hypoxia-induced mitochondrial oxidative stress and ER stress in the pathogenesis of preeclampsia/IUGR and discusses the potential therapeutic strategies targeting oxidative stress to treat both the pregnancy complications.
Pregnancy ; Female ; Humans ; Placenta ; Fetal Growth Retardation/etiology* ; Pre-Eclampsia/pathology* ; Reactive Oxygen Species ; Hypoxia/pathology* ; Pregnancy Complications/pathology* ; Endoplasmic Reticulum Stress

BACKGROUND AND PURPOSE: Neonatal hypoxic-ischemic encephalopathy (HIE) is one of the leading causes of neurological handicap in developing countries. Human umbilical cord blood (hUCB) CD34-positive (CD34⁺) stem cells exhibit the potential for neural repair. We tested the hypothesis that hUCB CD34⁺ stem cells and other cell types [leukocytes and nucleated red blood cells (NRBCs)] that are up-regulated during the acute stage of perinatal asphyxia (PA) could play a role in the early prediction of the occurrence, severity, and mortality of HIE. METHODS: This case-control pilot study investigated consecutive neonates exposed to PA. The hUCB CD34⁺ cell count in mononuclear layers was assayed using a flow cytometer. Twenty full-term neonates with PA and 25 healthy neonates were enrolled in the study. RESULTS: The absolute CD34⁺ cell count (p=0.02) and the relative CD34⁺ cell count (CD34+%) (p<0.001) in hUCB were higher in the HIE patients (n=20) than the healthy controls. The hUCB absolute CD34⁺ cell count (p=0.04), CD34⁺% (p<0.01), and Hobel risk scores (p=0.04) were higher in patients with moderate-to-severe HIE (n=9) than in those with mild HIE (n=11). The absolute CD34⁺ cell count was strongly correlated with CD34⁺% (p<0.001), Hobel risk score (p=0.04), total leukocyte count (TLC) (p<0.001), and NRBC count (p=0.01). CD34+% was correlated with TLC (p=0.02). CONCLUSIONS: hUCB CD34⁺ cells can be used to predict the occurrence, severity, and mortality of neonatal HIE after PA.
Asphyxia ; Case-Control Studies ; Cell Count ; Developing Countries ; Erythrocytes ; Fetal Blood* ; Humans* ; Hypoxia-Ischemia, Brain* ; Incidence* ; Infant, Newborn ; Leukocyte Count ; Mortality ; Pilot Projects* ; Stem Cells ; Umbilical Cord*

OBJECTIVETo study the effect of calcium-sensing receptor (CaSR) agonists and antagonists on the expression of CaSR in neonatal mice with persistent pulmonary hypertension (PPHN), and to clarify the role of CaSR in neonatal mice with PPHN.

METHODSForty-nine neonatal mice were randomly divided into four groups: control (n=10), hypoxia (PPHN; n=11), agonist (n=13), and antagonist (n=15). The mice in the PPHN, agonist, and antagonist groups were exposed to an oxygen concentration of 12%, and those in the control group were exposed to the air. The mice in the agonist and antagonist groups were intraperitoneally injected with gadolinium chloride (16 mg/kg) and NPS2390 (1 mg/kg) respectively once daily. Those in the PPHN and the control groups were given normal saline daily. All the mice were treated for 14 consecutive days. Hematoxylin and eosin staining and immunohistochemistry were used to observe the changes in pulmonary vessels. Laser confocal microscopy was used to observe the site of CaSR expression and measure its content in lung tissues. qRT-PCR and Western blot were used to measure the mRNA and protein expression of CaSR in lung tissues.

RESULTSCompared with the control group, the PPHN group had significant increases in the pulmonary small artery wall thickness and the ratio of right to left ventricular wall thickness (P<0.05), which suggested that the model was successfully prepared. Compared with the control group, the PPHN group had a significant increase in the mRNA and protein expression of CaSR (P<0.05), and the agonist group had a significantly greater increase (P<0.05); the antagonist group had a significant reduction in the mRNA and protein expression of CaSR (P<0.05).

CONCLUSIONSCaSR may play an important role in the development of PPHN induced by hypoxia in neonatal mice.

Animals ; Hypoxia ; complications ; Lung ; pathology ; Mice ; Myocardium ; pathology ; Persistent Fetal Circulation Syndrome ; etiology ; pathology ; Pulmonary Artery ; pathology ; RNA, Messenger ; analysis ; Receptors, Calcium-Sensing ; analysis ; genetics ; physiology

PURPOSE: The aim of this study is to prove the association between potential fetal hypoxia and retinopathy of prematurity (ROP) development and absolute nucleated red blood cell (aNRBC) is used to evaluate it in premature infants without any hypoxic or ischemic history. METHODS: Medical records of 43 premature infants with ROP who were admitted to the neonatal intensive care unit at Wonkwang University Hospital from January 2004 to December 2014 were analyzed retrospectively. We excluded 15 infants who had a confounding medical condition that could have increased the aNRBC count. Finally, 28 premature infants affected by ROP were enrolled and compared with 28 pair-matched controls. The aNRBC counts at birth in these infants were compared. Statistical analysis was performed with a paired t-test for continuous data, and a Fisher's exact test for categorical data. P<0.05 was considered significant. RESULTS: There were no significant differences in perinatal characteristics such as gestational age (GA), birth weight (BWt), Apgar scores, premature rupture of membrane (>24 hours), prenatal betamethasone, surfactant or respiratory distress syndrome between the ROP and the control infants. In addition, neither group differed in major morbidities such as patent ductus arteriosus, periventricular leukomalacia, intraventricular hemorrhage (> or =Grade 2), or bronchopulmonary dysplasia. Regardless of the severity of ROP, the aNRBC counts at birth in premature infants with ROP were not higher than in the control infants. CONCLUSION: The aNRBC counts at birth may not be related directly to the development of ROP.
Betamethasone ; Birth Weight ; Bronchopulmonary Dysplasia ; Ductus Arteriosus, Patent ; Erythrocyte Count* ; Erythrocytes* ; Fetal Hypoxia ; Gestational Age ; Hemorrhage ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Intensive Care, Neonatal ; Leukomalacia, Periventricular ; Medical Records ; Membranes ; Parturition ; Retinopathy of Prematurity* ; Retrospective Studies ; Rupture

PURPOSE: The purpose of this study was to investigate the clinical features of neonatal seizures and to identify prognostic factors of neurodevelopmental outcome in term infants who experienced clinical seizures. METHODS: A retrospective analysis was performed on 153full term and preterm infants with seizures from January 2008 to December 2013. Binary logistic regression analysis was applied to assess risk factors associated with neurological adverse outcomes using variables that were found to be significant on univariate analysis. RESULTS: During the study period, 102 (66.7%) term and 51 (33.3%) preterminfants were enrolled. The main cause of neonatal seizures was hypoxic ischemic encephalopathy (24.5%) in term infants and intracranial hemorrhage (74.5%) in preterm infants. The most common type of seizure was focal clonic seizure. Generalized tonic seizure was more commonly observed in preterm than in term infants. 39 out of 56 term infants with at least 12 months of neurologic follow-up showed normal outcomes while only one preterm infant showed normal development.Prognostic factors related to adverse neurodevelopmental outcomes in term infants were perinatal history of fetal distress, etiology of hypoxic ischemic encephalopathy, severity of EEG(Electroencephalogram) abnormality, evidence of hypoxic ischemic encephalopathy on brain magnetic resonance imaging, and the need for multiple antiepileptic drugs for seizure control. CONCLUSION: Preterm infants showed poorer neurodevelopmental outcomes compared to term infants. The etiology of seizures, treatment response, neuroimaging and electroencephalographic findings were important in predicting the developmental outcome in term infants with seizures.
Anticonvulsants ; Brain ; Epilepsy, Partial, Motor ; Fetal Distress ; Follow-Up Studies ; Humans ; Hypoxia-Ischemia, Brain ; Infant ; Infant, Newborn ; Infant, Premature ; Intracranial Hemorrhages ; Logistic Models ; Magnetic Resonance Imaging ; Neuroimaging ; Prognosis ; Retrospective Studies ; Risk Factors ; Seizures*

OBJECTIVETo explore the effect of human umbilical cord blood mononuclear cells (UCBMC) promoting nerve behavior function and brain tissue recovery of neonatal SD rat with hypoxic ischemic brain injury (HIBI).

METHODA modified newborn rat model that had a combined hypoxic and ischemic brain injury as described by Rice-Vannucci was used, early nervous reflex, the Morris water maze and walking track analysis were used to evaluate nervous behavioral function, and brain MRI, HE staining to evaluate brain damage recovery.

RESULTNewborn rat Rice-Vannucci model showed significant brain atrophy, obvious hemiplegia of contralateral limbs,e.g right step length [(7.67 ± 0.46) cm vs. (8.22 ± 0.50) cm, F = 1.494] and toe distance [(0.93 ± 0.06) cm vs. (1.12 ± 0.55) cm, F = 0.186] were significantly reduced compared with left side, learning and memory ability was significantly impaired compared with normal control group (P < 0.01); Cliff aversion [(8.44 ± 2.38) s vs.(14.22 ± 5.07) s, t = 4.618] and negative geotaxis reflex time [(7.26 ± 2.00) s vs. (11.76 ± 3.73) s, t = 4.755] on postnatal 14 days of HIBI+ transplantation group were significantly reduced compared with HIBI+NaCl group (P < 0.01) ; the Morris water maze experiment showed escape latency [ (23.11 ± 6.64) s vs. (34.04 ± 12.95) s, t = 3.356] and swimming distance [ (9.12 ± 1.21) cm vs.(12.70 ± 1.53) cm, t = 17.095] of HIBI+transplantation group were significantly reduced compared with those of HIBI+NaCl group (P < 0.01) ; the residual brain volume on postnatal 10 d [ (75.37 ± 4.53)% vs. (67.17 ± 4.08)%, t = -6.017] and 67 d [ (69.05 ± 3.58)% vs.(60.83 ± 3.69)%, t = -7.148]of HIBI+ transplantation group were significantly larger than those of HIBI+NaCl group (P < 0.01); After human UCBMC transplantation, left cortical edema significantly reduced and nerve cell necrosis of HIBI+ transplantation group is not obvious compared with HIBI+NaCl group.

CONCLUSIONHuman UCBMC intraperitoneal transplantation significantly promoted recovery of injured brain cells and neurobehavioral function development.

Animals ; Animals, Newborn ; Atrophy ; etiology ; pathology ; Brain ; diagnostic imaging ; pathology ; Cerebral Cortex ; pathology ; Cord Blood Stem Cell Transplantation ; methods ; Disease Models, Animal ; Female ; Fetal Blood ; cytology ; Humans ; Hypoxia-Ischemia, Brain ; complications ; pathology ; therapy ; Learning Disorders ; etiology ; prevention & control ; Leukocytes, Mononuclear ; cytology ; transplantation ; Magnetic Resonance Imaging ; Male ; Maze Learning ; Neurons ; pathology ; Psychomotor Performance ; Radiography ; Rats ; Rats, Sprague-Dawley ; Transplantation, Heterologous

OBJECTIVETo study the effects of umbilical cord blood monocytes (UCBMC) transplantation on erythropoietin (EPO) protein and oligodendrocyte progenitor cells in hypoxia-ischemia (HI) neonatal rats.

METHODSForty seven-day-old Sprague-Dawley rats were randomly divided into normal control (N), HI, UCBMC and HI+UCBMC groups (n=10 each). Hypoxic-ischemic brain damage (HIBD) model was prepared according to the Rice method. Twenty-four hours after hypoxia, the N and HI groups were injected with 2 μL phosphate buffered saline (PBS), and the UCBMC and HI+UCBMC groups were injected with 3×10(6) UCBMC via the lateral ventricle. EPO protein and oligodendrocyte progenitor cells in the subventricular zone of the injured brain were observed by EPO/DAPI and NG2/DAPI immunofluorescence double staining, and their correlation was analyzed.

RESULTSSeven days after transplantation, there were more NG2(+)DAPI(+) and EPO(+)DAPI(+) cells in the HI+UCBMC group than in the UCBMC (P<0.05), N and HI groups (P<0.01). More NG2(+)DAPI(+) and EPO(+)DAPI(+) cells were observed in the UCBMC group compared with the N and HI groups (P<0.01). There were more NG2(+)DAPI(+) cells in the N group than in the HI group (P<0.01). The number of NG2(+)DAPI(+) cells was correlated with the number of EPO(+)DAPI(+) cells in the HI+UCBMC group (r=0.898, β=1.4604, P<0.01).

CONCLUSIONSUCBMC can promote expression of oligodendrocyte progenitor cells, which is correlated with an increase in EPO protein and thus repairs brain white matter damage in neonatal rats with HIBD.

Animals ; Animals, Newborn ; Erythropoietin ; analysis ; biosynthesis ; Fetal Blood ; cytology ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; therapy ; Monocytes ; transplantation ; Oligodendroglia ; pathology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; pathology

The vasculosyncytial membrane (VSM), primary site of fetomaternal exchange is formed when syncytiotrophoblast surrounds the terminal villi and make a close contact with capillaries. Some syncytiotrophoblast forms thin single layer of villous and some syncytial nuclei become piled up to form the syncytial knots (SKs). Undoubtedly there is a clear-cut inverse relation between villous VSM and fetal hypoxia. In preeclampsia (PE) the hypoxia injury disrupts the syncytial architecture which in turn initiates other complications of PE. Present study was designed to observe the morphological and histomorphometric features of 84 placentas from control and PE (42 each) collected from Department of Obstetrics and Gynecology. Neonatal weight and placental weight were reduced in PE than the controls but the feto-placental index did not differ. The SK density and VSM thickness was found to be increased and was statistically significant in PE cases. In relation to SKs, the VSM thickness was twofold increased than the controls and was statistically significant. The SKs in the present study were classified as type-1, 2a, 2b, and 3. Type 1 was found to be 62% in control and 47% in PE, type 2a and 2b were 38% in control and 37% in PE, and type 3 was in 8% of PE cases. All the parameters of present study reveal the adverse effects of PE influencing on both morphological and microscopical features of the placenta resulting in fetal hypoxia.
Anoxia ; Capillaries ; Fetal Hypoxia ; Gynecology ; Membranes ; Obstetrics ; Placenta ; Pre-Eclampsia ; Trophoblasts

The vasculosyncytial membrane (VSM), primary site of fetomaternal exchange is formed when syncytiotrophoblast surrounds the terminal villi and make a close contact with capillaries. Some syncytiotrophoblast forms thin single layer of villous and some syncytial nuclei become piled up to form the syncytial knots (SKs). Undoubtedly there is a clear-cut inverse relation between villous VSM and fetal hypoxia. In preeclampsia (PE) the hypoxia injury disrupts the syncytial architecture which in turn initiates other complications of PE. Present study was designed to observe the morphological and histomorphometric features of 84 placentas from control and PE (42 each) collected from Department of Obstetrics and Gynecology. Neonatal weight and placental weight were reduced in PE than the controls but the feto-placental index did not differ. The SK density and VSM thickness was found to be increased and was statistically significant in PE cases. In relation to SKs, the VSM thickness was twofold increased than the controls and was statistically significant. The SKs in the present study were classified as type-1, 2a, 2b, and 3. Type 1 was found to be 62% in control and 47% in PE, type 2a and 2b were 38% in control and 37% in PE, and type 3 was in 8% of PE cases. All the parameters of present study reveal the adverse effects of PE influencing on both morphological and microscopical features of the placenta resulting in fetal hypoxia.
Anoxia ; Capillaries ; Fetal Hypoxia ; Gynecology ; Membranes ; Obstetrics ; Placenta ; Pre-Eclampsia ; Trophoblasts