Natural killer cells 56bnght16- have higher counts in the umbilical cord blood than in the adult peripheral blood

ABSTRACT Introduction and hypothesis: Umbilical cord blood (UCB) is an alternative source of hematopoietic stem cells for allogeneic hematopoietic stem cell transplantation in the absence of a compatible donor. The UCB transplantation has a lower incidence of chronic graft versus host disease (GvHD), but is associated with slower engraftment and slower immune reconstitution, compared to other sources. Dendritic cells (DCs) and Natural Killer cells (NKs) play a central role in the development of GvHD and the graft versus leukemia (GvL) effect, as well as in the control of infectious complications. Method: We quantified by multiparametric flow cytometry monocytes, lymphocytes, NK cells, and DCs, including their subsets, in UCB samples from 54 healthy newborns and peripheral blood (PB) from 25 healthy adult volunteers. Results: In the UCB samples, there were higher counts of NK cells 56bright16- (median 0.024 × 109/L), compared to the PB samples (0.012 × 109/L, p < 0.0001), NK 56dim16bright (median 0.446 × 109/L vs. 0.259 × 109/L for PB samples, p = 0.001) and plasmacytoid dendritic cells (pDCs, median 0.008 × 109/L for UCB samples vs. 0.006 × 109/L for PB samples, p = 0.03). Moreover, non-classic monocyte counts were lower in UCB than in PB (median 0.024 × 109/L vs. 0.051 × 109/L, respectively, p < 0.0001). Conclusion: In conclusion, there were higher counts of NK cells and pDCs and lower counts of non-classic monocytes in UCB than in PB from healthy individuals. These findings might explain the lower incidence and severity of chronic GvHD, although maintaining the GvL effect, in UCB transplant recipients, compared to other stem cell sources.

ASSOCIATION OF SERUM FERRITIN IN POST COVID-19 ANAEMIC PATIENTS TREATED WITH UMBILICAL CORD BLOOD AND HAEMATINICS.

COVID-19 patients commonly present with lower respiratory symptoms with other systemic involvement. Haematological manifestation such as low haemoglobin, thrombocytopenia, lymphocytopenia also common in COVID19 patients. In this study, we investigated prevalence, association with serum ferritin in post COVID-19 anaemic patients, after human umbilical cord blood transfusion in relation to control group. Among 155 COVID-19 RT-PCR positive patients 36 (23%) was anaemic. In our study 18 patients was transfused human umbilical cord blood, 12 patients were treated with haematinics and 6 patients denied taking any of the above. In most cases anaemia was moderate to severe that may be due to inflammation or due to pre-existing iron deficiency.Umbilical cord blood transfusion to post COVID -19 patients for the treatment of anaemia because of the unique composition of UCB. Haematological analysis and serum ferritin estimation reflecting the treatment out come in post COVID-19 anaemic patients. There was a difference between the dependent variable's serum ferritin (p <.001) in anaemic COVID-19 patients. In conclusion, our result highlight serum ferritin is widely used in diagnosis and monitoring of COVID-19 disease.

Treatment of severe aplastic anemia and hematopoietic stem cell transplantation / 器官移植

Severe aplastic anemia (SAA) is a severe bone marrow failure syndrome caused by multiple causes, which is clinically manifested with severe anemia, infection and bleeding. The complex pathogenesis of SAA has not been fully understood. SAA is characterized with acute onset, severe disease condition and rapid progression. At present, with the in-depth study of SAA and the improvement of diagnosis and treatment, the therapeutic strategy for SAA has been evolved from classical immunosuppressive therapy based on antithymocyte globulin and cyclosporine to the application of thrombopoietin receptor agonist and combined treatment based on allogeneic hematopoietic stem cell transplantation, which may promote the reconstruction of hematopoietic function of SAA patients to varying degree and significantly improve survival and clinical prognosis, becoming the research hotspot of SAA treatment. In this article, new advances in the treatment of SAA at home and abroad were reviewed.

Association between 25-hydroxyvitamin D levels and incidence of allergy and infection in infants aged 0–6 months

@#Introduction: Vitamin D levels are known to be related to prevalence of allergy and infection in children. However, vitamin D levels in infants’ umbilical cord blood need to be investigated. Therefore, this study aimed to determine association between 25-hydroxyvitamin D [25(OH)D] levels and incidence of allergy and infection in children. Methods: A longitudinal study involving 38 full-term newborns was conducted. Serum 25(OH)D levels in infants’ umbilical cord and venous blood were measured at birth and six months, respectively. 25(OH)D levels were classified as insufficient (<20 ng/mL) and sufficient (>20 ng/mL). Parents filled out questionnaires about their children’s allergy and infection symptoms. Paired t-test was performed to compare the 25(OH)D levels at birth and at six months. Chisquared test was conducted to determine relationship between 25(OH)D levels and incidence of infection and allergy in children. Results: 25(OH)D levels in venous blood of 6-month-old infants were significantly higher than in umbilical cord blood (50.44±13.59 ng/mL vs. 20.70±6.60 ng/mL, p<0.001). In addition, 25(OH)D level insufficiency in umbilical cord blood was associated with infection (p<0.05). However, there was no incidence of allergy, and exclusive breastfeeding and sun exposure were not associated with vitamin D levels in 6-month-old infants. Conclusion: We conclude that 25(OH)D level insufficiency in umbilical cord blood was associated with incidence of infection in the first six months of life.

Analysis of pathogenic microorganisms in 64 077 cases of umbilical cord blood / 中国输血杂志

【Objective】 To investigate the detection of pathogenic microorganisms in umbilical cord blood and maternal blood from 2012 to 2021, so as to improve the collection of umbilical cord blood and guarantee the safety of umbilical cord blood hematopoietic stem cells (HSC) . 【Methods】 Detection results of pathogenic microorganisms of umbilical cord blood and maternal blood among 64 077 cases from Tianjin Cord Blood Bank from 2012 to 2021 were retrospectively analyzed. 【Results】 A total of, 2 072 cases (3.23%) were detected positive, among which, 184 cases (0.29%) were positive for aerobic bacteria culture, 1 504 cases (2.34%) were positive for anaerobic bacteria culture, and 384 cases (0.60%) were positive for both aerobic and anaerobic bacteria culture. From 2012 to 2021,the overall positive rate showed a downward trend, with a difference in the positive rate between each year (P<0.05). The positive rate of anaerobic bacteria was higher than that of aerobic bacteria and that of anaerobic and aerobic bacteria (P<0.05). After Gram staining, the microscopic detection rate of bacterial positive samples was highest in G- bacilli, followed by G+ bacilli, G+ cocci, G- cocci and others. Among the 64 077 cases, 169 cases (0.26%) showed reactivity in cord blood tests and 1 231 cases (1.92%) showed reactivity in maternal blood tests. Umbilical cord blood and maternal blood HIV-Ag/Ab tests showed reactivity after initial screening. After confirmation by Western blotting, there was 1 case of uncertain maternal blood, while the rest were negative. The reactive rates of anti-TP (0.12%) and anti- HCV (0.11%) in umbilical cord blood were higher than those of HBsAg (0.03%) and CMV-IgM (1/64 077).There was a difference in the reactive rate of anti-TP detection in umbilical cord blood between different years (P<0.05),while there was no statistically significant difference in that of HBsAg, anti-HCV and CMV-IgM (P> 0.05).The reactive rate of HBsAg in maternal blood (1.38%) was higher than that of CMV-IgM(0.29%) , anti-TP(0.13%) and anti-HCV (0.12%) . There were differences in the reactive rates of HBsAg, anti-HCV ,and anti-TP in maternal blood among different years (P<0.05),and that of HBsAg showed a decreasing trend, while the reactive rate of CMV-IgM was not statistically significant (P>0.05). The reactive rates of HBsAg and CMV-IgM detected in maternal blood were significantly higher than those in umbilical cord blood (P<0.05) . The reactive rates of anti-HCV and anti-TP in maternal blood were consistent with those in umbilical cord blood (P>0.05). 【Conclusion】 The reactive rates of anti-HIV and CMV-IgM in cord blood, and that of anti-HIV in maternal blood are low, but those of anti-TP and anti-HCV in cord blood are relatively high. The reactive rate of HBsAg is high in maternal blood,but with a downward trend,but low in umbilical cord blood due to maternal-infantile transmission blocking. The detection of transfusion transmitted pathogens and bacteria plays a critical role on the safety of umbilical cord blood HSCs. Effective detection of transfusion transmitted pathogens and culture of bacteria are the key to ensure the quality of umbilical cord blood, which can improve the safety of umbilical cord blood HSCs transplantation.

Expansion of cord blood stem cells in fibronectin-coated microfluidic bioreactor

ABSTRACT Background: Hematopoietic stem/progenitor cell transplantation is the main treatment option for hematological malignancies and disorders. One strategy to solve the problem of low stem cell doses used in transplantation is pre-transplant expansion. We hypothesized that using fibronectin-coated microfluidic channels would expand HSPCs and keep self-renewal potential in a three-dimensional environment, compared to the conventional method. We also compared stem cell homing factors expression in microfluidic to conventional cultures. Materials and methods: A microfluidic device was created and characterized by scanning electron microscopy. The CD133+ cells were collected from cord blood and purified. They were subsequently cultured in 24-well plates and microfluidic bioreactor systems using the StemSpan serum-free medium. Eventually, we analyzed cell surface expression levels of the CXCR4 molecule and CXCR4 mRNA expression in CD133+ cells cultured in different systems. Results: The expansion results showed significant improvement in CD133+ cell expansion in the microfluidic system than the conventional method. The median expression of the CXCR4 in the expanded cell was lower in the conventional system than in the microfluidic system. The CXCR4 gene expression up-regulated in the microfluidic system. Conclusion: Utilizing microfluidic systems to expand desired cells effectively is the next step in cell culture. Comparative gene expression profiling provides a glimpse of the effects of culture microenvironments on the genetic program of HSCs grown in different systems.

Effect of hypertension disorders during pregnancy on umbilical cord blood hemoglobin, serum ferritin at birth and outcome in premature newborns / 中华实用儿科临床杂志

Objective:To investigate the effects of hypertensive disorders during pregnancy (HDP) on preterm newborns in terms of umbilical cord blood serum ferritin (SF), hemoglobin (Hb) at birth and outcome.Methods:Among inpatients of the First Maternity and Infant Health Hospital Affiliated to Tongji University from October 1, 2015 to December 31, 2016, totally 1 419 cases of preterm newborns were prospectively collected.Preterm infants whose mothers with diagnosis of HDP were recruited as the HDP group.Meanwhile, premature newborns whose mothers without HDP were recruited as the control group.Umbilical cord blood SF levels, Hb levels at birth, outcome of preterm newborns and the basic information for maternity were compared between the two groups.The data of normal distribution between the two groups were compared by independent sample t test.The count data was tested by χ2, and the count data with frequency <5 was tested by Fisher′ s exact test. Results:SF levels of HDP group were significantly lower than the control group [(85.6±67.2) μg/L vs. (103.9±95.5) μg/L]. But Hb levels of HDP group were much higher than the control group [(206.2±33.8) g/L vs. (193.2±31.9) g/L]. The difference between two groups was statistically significant ( t=2.791, 4.825 all P<0.05). Umbilical cord blood SF levels were negatively correlated with Hb levels at birth ( r=-0.120, P<0.001). Moreover, compared to the control group, statistically significant lower incidence of neonatal respiratory distress syndrome (NRDS), pneumonia and bronchopulmonary dysplasia (BPD) in HDP group was observed (all P<0.05). Conclusions:HDP was correlated with umbilical cord blood SF levels and Hb levels at birth in premature newborns.Higher Hb levels and relatively lower incidences of NRDS, pneumonia and BPD were observed in these newborns delivered by mothers with diagnosis of HDP.

Case report of unrelated cord blood transplantation for the treatment of recurrent refractory Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis with intestinal perforation and literature review / 中华实用儿科临床杂志

Objective:To examine the clinical experience and efficacy of unrelated cord blood transplantation (UCBT) in the treatment of recurrent refractory Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH) in children.Methods:The clinical data of a patient with recurrent refractory EBV-HLH and intestinal perforation who was treated by UCBT in Department of Pediatrics, the First Affiliated Hospital of Zhengzhou University in September 2015 and finally cured were retrospectively analyzed.Meanwhile, literature was reviewed.Results:The patient, male, 1 year and 6 months, was admitted to the hospital with " fever for 15 days, rash for 9 days" as the main complaint, mainly manifested as high fever, large liver, spleen, lymph nodes, rapidly progressing pancytopenia, liver function damage, phagocytic blood cells on bone marrow smear, diagnosed as EBV-HLH in September 2015.The patient received chemotherapy according to the HLH-2004 protocol developed by the International Association of Cell Societies.During the treatment, he suffered two recurrence during the maintenance period, and a second-line rescue treatment was adopted, namely, " Pegaspargase, Doxorubicin liposome, Etoposide and Methylprednisolone" (L-DEP regimen) chemotherapy.The complete relief of diagnostic indexes for hemophagocytic lymphohistiocytosis was evaluated after chemotherapy.The patient developed sudden intestinal perforation and underwent emergency surgical surgery, enteroenterostomy.After the condition was stabilized, the patient was pretreated with the " Fludarabine+ Busulfan+ Cyclophosphamide" (Flu+ BU+ CY) therapy and then treated with UCBT, with intravenous nutritional support provided during the entire process.Neutrophil and platelet implantation was implemented on day 13 and day 35 after transplantation, respectively.The chimeric rate was 100%, and the implantation was a success.Hepatic veno-occlusive disease, fungal pneumonia and skin graft-versus-host disease (GVHD) Ⅱ occurred on the 15 th day, 22 nd day and 26 th day after transplantation, respectively.The corresponding symptoms improved after treatment.On day 49 after transplantation, phase Ⅱ " enterostomy fistula" was performed.The patient was followed up to 70 months after transplantation, and generally in good condition.His symptoms relieved, and no chronic GVHD and other comorbidities occurred. Conclusions:Allogeneic hematopoietic stem cell transplantation is the only possible effective means of treating relapsed refractory EBV-HLH in children.In the absence of a suitable sibling or unrelated donor, unrelated cord blood stem cells can be used as a graft source.Enterostomy after intestinal perforation is not contraindicated for transplantation.

Optimización de unidades formadoras de colonias y detección del virus de papiloma humano en sangre de cordón umbilical / Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Resumen Introducción: Se requiere analizar diversos parámetros para el control de calidad adecuado de las unidades de sangre de cordón umbilical (USCU) cuando se utilizan con fines terapéuticos. Objetivo: Optimizar las unidades formadoras de colonias (UFC) de cultivos clonogénicos y detectar el genoma del virus del papiloma humano (VPH) en USCU. Métodos: Se incluyeron 141 muestras de sangre de cordón umbilical (SCU), de segmento y de UFC de cultivos clonogénicos de USCU. Se realizó extracción de ADN, cuantificación y amplificación por PCR del gen endógeno GAPDH. Se detectó el gen L1 del VPH con los oligonucleótidos MY09/MY11 y GP5/GP6+; los productos de PCR se migraron en electroforesis de agarosa. El ADN purificado de las UFC se analizó mediante electroforesis de agarosa y algunos ADN, con la técnica sequence specific priming. Resultados: La concentración de ADN extraído de UFC fue superior comparada con la de SCU (p = 0.0041) y la de segmento (p < 0.0001); así como la de SCU comparada con la de segmento (p < 0.0001). Todas las muestras fueron positivas para la amplificación de GAPDH y negativas para MY09/MY11 y GP5/GP6+. Conclusiones: Las USCU criopreservadas fueron VPH netativas; además, es factible obtener ADN en altas concentraciones y con alta pureza a partir de UFC de los cultivos clonogénicos.

Abstract Introduction: Analysis of several markers is required for adequate quality control in umbilical cord blood units (UCBU) when are used for therapeutic purposes. Objective: To optimize colony-forming units (CFU) from clonogenic cultures and to detect the human papillomavirus (HPV) genome in UCBU. Methods: One hundred and forty-one umbilical cord blood (UCB), segment or CFU samples from UCBU clonogenic cultures were included. DNA extraction, quantification and endogenous GAPDH gene PCR amplification were carried out. Subsequently, HPV L1 gene was detected using the MY09/MY11 and GP5/GP6+ oligonucleotides. PCR products were analyzed with electrophoresis in agarose gel. CFU-extracted purified DNA was analyzed by electrophoresis in agarose gel, as well as some DNAs, using the SSP technique. Results: CFU-extracted DNA concentration was higher in comparison with that of UCB (p = 0.0041) and that of the segment (p < 0.0001), as well as that of UCB in comparison with that of the segment (p < 0.0001). All samples were positive for GAPDH amplification and negative for MY09/MY11 and GP5/GP6+. Conclusions: Cryopreserved UCBUs were HPV-negative. Obtaining CFU DNA from clonogenic cultures with high concentrations and purity is feasible.

Comparison of immune reconstitution after peripheral blood stem cell transplantation and umbilical cord blood transplantation / 中华检验医学杂志

Objective:To explore the differences of immune reconstitution between peripheral blood stem cell transplantation and umbilical cord blood transplantation.Methods:A total of 300 patients (aged 18 (8, 33), 163 males and 137 females) with malignant hematological diseases who received allogeneic hematopoietic stem cell transplantation in the First Affiliated Hospital of University of Science and Technology of China from January 2018 to March 2020 were enrolled in this study, including 255 cases of umbilical cord blood transplantation and 45 cases of peripheral blood stem cell transplantation. Multicolor flow cytometry was applied to analyze lymphocyte subsets of the percentages and absolute counts in the two donor types and peripheral blood of patients after receiving hematopoietic stem cell transplantation. The differences between the two grafts were compared, and the lymphocyte subsets results were evaluated at 1, 2, 3, 4, 6, 9, 12, and 18 months after transplantation. 18-month disease-free survival (DFS) within the 300 patients under the two transplantation types were retrospectively analyzed.Results:1. The proportion of NKT cells in peripheral blood group was significantly higher than that in cord blood group (2.79% vs 0.24%, P<0.001). 2. The proportion of helper T cells in the UCBT group was higher than that in the PBSCT group, as well as the counts 6 months after transplantation ( P<0.05). 3. The proportion of NK1 cells (3 rd to 9 th month) and count (4 th to 12 th month) in UCBT group were significantly higher than those in PBSCT group ( P<0.05). 4. NKT cells in the UCBT group were lower than those in the PBSCT group (proportion and count) throughout the monitoring process ( P≤0.001). 5. The proportion of DNT cells (within 1 year) and count (within 6 months) in the UCBT group were significantly lower than those in the PBSCT group ( P<0.05). Conclusions:Compared with the peripheral blood stem cell transplantation group, the umbilical cord blood transplantation patients had a faster rate of lymphocyte reconstitution, and patients received umbilical cord blood transplantation had a stronger ability of immune reconstitution and could achieve long-term hematopoiesis.

Efect of PolyCHb on enhancing tissue oxygen supply and the treatment of remvatinib for liver cancer graft: Preliminary observation / 中国输血杂志

【Objective】 To investigate the effect of Polymerized human cord hemoglobin (PolyCHb) on the sensitivity of hepatocellular carcinoma grafts to lumvalatinib in nude mice. 【Methods】 Hep3B hepatoma cells were subcutaneously transplanted in 18 nude mice to establish tumor graft model. Mice were randomly divided into 3 groups: control group (the saline 90 mg·kg-1·d-1), monotherapy group (Lenvatinib10 mg·kg-1·d-1), and sensitized group (Lenvatinib mg·kg-1·d-1, polyCHB 600 mg/kg twice a week) for 28 days. The tumor volume was measured regularly and the growth curve was drawn. On day 29, the nude mice were sacrificed, the tumor was stripped and weighed, and the pathomorphological differences of each group were evaluated by HE section staining. The expression levels of hypoxia-inducing factor (HIF-1α), CD34, VEGF, CD44, MMP-9, and Glut-1 in tumor tissues of each group were determined by immunohistochemistry. The content of reactive oxygen species (ROS) in tumor tissues of each group was determined by dihydroethyl ingot method. 【Results】 The tumor growth rate and tumor volume in the sensitized group decreased significantly compared with the control group and the solo drug group. On day 29, the tumor volumes of the control group, the monotherapy group and the sensitization group were (2 076.46±350.25)mm3, (1 035.96±84.16)mm3 and (892.66±104.46)mm3, respectively. Tumor weight was (1.61±0.52)g, (0.45±0.10)g, and (0.34±0.13)g, respectively. Immunohistochemical score of HIF-1α was 75±23 vs 45±18 vs 18±11, VEGF was 52±8 vs 67±16 vs 35±4, CD34 was 40±7 vs 50±13 vs 28±7, CD44 was 37±15 vs 30±7 vs 15±3, Glut-1 was 74±41 vs 51±30 vs 14±18, MMP-9 was 51±7 vs 62±20 vs 33±3, respectively(P<0.05). The malignant degree of the sensitized group was decreased by HE section staining, which was significantly lower than that of the solo drug group and the control. The ROS content in the sensitized group was higher than that in the solo drug group and the control. 【Conclusion】 PolyCHb can reduce the expression of HIF-1α and its downstream pathway related molecules by increasing oxygenation of hepatocellular carcinoma tissues in nude mice, delay tumor growth and reduce tumor volume in a certain period, thus increase the therapeutic effect of lenvalatinib on hepatocellular carcinoma grafts in tumor bearing nude mice models.

Clinical study on haploidentical hematopoietic stem cell transplantation for children with bone marrow failure syndromes / 中华实用儿科临床杂志

Objective:To explore the efficacy of haploidentical hematopoietic stem cell transplantation (haplo-HSC) combined with tpCB in the treatment of children with bone marrow failure syndromes (BMFs).Methods:The clinical chara-cteristics of 78 BMFs pediatric patients, including inherited BMFs (4 cases) and acquired BMFs (74 cases) under-went haplo-HSC combined with the third-party cord blood (tpCB) in Chinese People′s Liberation Army General Hospital-Sixth Medical Center between July 2012 and July 2019 and were retrospectively analyzed, with 41 males and 37 females.Among them, 73 cases experienced first transplantation and 5 cases accepted second transplantation, with the median age of 5.6 years.The conditioning regimen was based on Busulfan, with 74 acquired BMFs cases using non-myeloablative and the remaining 4 cases using myeloablative.The prophylaxis of acute graft versus host disease (aGVHD) includes Cyclosporine, Mycophenolate mofetil (MMF) and Methotrexate.All patients received bone marrow from haploid donor and tpCB on day 1 and peripheral stem cell from haploid donor on day 2.The median dose of the total donor nucleated cells was 12.19×10 8/kg of recipient weight and CD 34+ cell dose was 6.13×10 6/kg of recipient weight. Results:The median time of granulocytes over 0.5×10 9/L and platelets over 20×10 9/L were + 13 d and + 17 d, respectively.All patients displayed complete donor-type chimerism at + 30 d. No primary graft failure occurred in any patient and second graft failure occurred in two cases.The incidence rate of grade ⅡtoⅣ and grade Ⅲ to Ⅳ aGVHD were 39.0% and 13.9%, respectively.The incidence of chronic GVHD with limited type and extensive type were 7.8% (95% CI: 7.1%-8.5%) and 2.6% (95% CI: 2.1%-3.1%), respectively.the median follow-up was 1 550 days, and 76 patients survived with free disease.The rate of transplant related mortality was 2.8%, and both of the estimated 5-year overall survival and failure-free survival rate were 97.2%(95% CI: 96.8%-97.6%). Conclusions:Haplo-HSC and umbilical cord blood can quickly provide hematopoietic stem cells.The results of haplo-HSC combined with the tpCB in pediatric patients with life-threatening BMFs are promising.

Study of relationship between umbilical cord blood hemoglobin percentage and perinatal asphyxia

Background: Perinatal asphyxia may be caused by perinatal anemia. The pathophysiology and neurodevelopment effects are theoretically different from other causes of fetal asphyxia. Severe asphyxia can occur in infants around the time of birth by various reasons. The aim of this study to find the relationship between cord blood hemoglobin and perinatal asphyxia.Methods: This was a retrospective comparative study in department of OBG In tertiary care health centre. Umbilical cord blood samples were collected from 100 newborns with asphyxia at birth as study group and 100 newborns with non asphyxia as control group. Hemoglobin was measured colorimetrically.Results: This study finds that maximum number of patients in both the control and study group had hemoglobin in the range of 16.3-17.3 gm/dl. The difference was not statistically significant. P value>0.05.Conclusions: Hematological changes observed early after delivery can determine the duration of hypoxemia (acute versus chronic) Perinatal anemia causing moderate to severe perinatal asphyxia is associated with a higher risk for neonatal mortality. All survivors with perinatal anemia, however, showed no abnormalities in neurodevelopment in contrast to children who were born asphyxiated due to various another causes. The underlying pathophysiological mechanism for the favorable NDO in the perinatal anemia group needs further elucidation.

Study of intrapartum fetal distress with the help of cardiotocography and its correlation with umbilical cord blood sampling

Background: Even in low risk mothers, fetal acidosis occurs as in high risk groups. Aim of fetal monitoring is to detect early response to intrauterine hypoxia and prevent irreversible neurological damage and death. Objective of this study was to correlate the intrapartum fetal distress with the help of cardiotocography CTG with umbilical cord blood sampling.Methods: A total 100 consecutive patients attending the labor ward were studied. Immediately at birth, before the baby’s first breath and before delivery of the placenta, the umbilical cord blood was collected as per the standard guidelines laid down in the standard textbooks. Fetal acidosis was assessed by umbilical cord arterial blood pH. Fetal acidosis was considered when umbilical artery pH <7.2. Cardiotocography features were used to clinically diagnose fetal distress.Results: Most of the mothers were multigravida. They belonged to the age group of 20-25 years. Only 18% had abnormal CTG. Out of 50 mothers with normal vaginal delivery, all had normal CTG. Out of 43 mothers who were delivered by LSCS, no one had normal CTG, 25 had indeterminate CTG and 18 had abnormal CTG. As CTG became abnormal, proportion of mothers with the thick meconium increased. NICU admission proportion increased as CTG changed from normal to the abnormal. There was a significant association between the abnormal CTG and the umbilical cord blood pH being acidic.Conclusions: CTG is a simple test, easy to perform and can alert obstetrician for necessary interventions in case of an abnormal CTG. It can detect fetal distress in labor thus helping to reduce neonatal morbidity by early intervention in cases of abnormal tracing.

A Clinical Study for Prediction of Perinatal Asphyxia using Nucleated Red Blood Cells as an Indicator in Umbilical Cord Blood of Newborns

Introduction: Perinatal asphyxia has become the leadingcause of death for newborns. Since the parameters that arebeing routinely used as a predictor for perinatal asphyxia suchas thick meconium stained liquor, non-reassuring fetal heartpatterns, low Apgar scores, fetal or cord blood pH do notshow consistent correlation with fetal acidosis but nucleatedred blood cell counts in umbilical venous blood of neonateshas been reported as a possible marker of perinatal asphyxia.The number of nucleated red blood cells (nRBCs)/100 whiteblood cells is variable but is rarely greater than 10 in normalneonates. The aim of this study was to study the using ofnucleated red blood cells in umbilical cord blood of newbornsas an indicator for perinatal asphyxia.Material and methods: The study comprised of intramuralterm neonates with and without asphyxia. It was a casecontrol study conducted in the Department of Pediatrics incollaboration with Department of Obstetrics and Gynaecology,Rohilkhand Medical College and hospital, Bareilly fromNovember 2017 to October 2018. There were 50 asphyxiatednewborns and 50 healthy newborns in the study. The meanlevels of nRBCs in cord blood were significantly higher(p value<0.001) in the asphyxial group (54.06+-22.42) ascompared to control group (10.32+-5.86).Results: Nucleated red blood cells were found to be stronglyassociated with perinatal asphyxia. The mean values of cordblood nRBCs in cases were 54.06+-22.42 and in controlgroup were 10.32+-5.86. The p value being <0.001 which wasstatistically significant indicating the correlation.Conclusions: Therefore it was concluded that nucleated redblood cells in umbilical cord blood of newborns can be used asan effective test for prediction of perinatal asphyxia.

Supporting effect of human skeletal muscle-derived myoendothelial cells on hematopoietic stem/progenitor cells in vitro / 中国组织工程研究

BACKGROUND: Human skeletal muscle derived myoendothelial cells (MECs) are located in the vascular wall and co-express the markers of muscle stem cells and vascular endothelial cells (CD56+CD34+CD144+CD45-). Studies have shown that MECs are similar to mesenchymal stem cells, express the surface markers of mesenchymal stem cells and have the potential of multidirectional differentiation. OBJECTIVE: To establish an in vitro culture system for human umbilical cord blood CD34+ cells with MECs as trophoblastic layer, and to evaluate the in vitro supporting effect of human skeletal muscle MECs on hematopoietic stem/progenitor cells by measuring the changes in the number, immunophenotype and colony forming ability of CD34+ cells before and after culture. METHODS: There were three groups in the experiment. In experimental group, human umbilical cord blood CD34+ cells were co-cultured with MECs as the nourishing layer; in control group, human umbilical cord blood CD34+ cells were co-cultured with bone marrow mesenchymal stem cells as the nourishing layer; and in blank control group, human umbilical cord blood CD34+ cells were cultured alone without the nourishing layer. The main outcome measures, including the number of human umbilical cord blood CD34+ cells, immunophenotype of blood cells and colony formation ability of hematopoietic stem/progenitor cells were analyzed and compared at 1, 2, and 4 weeks after co-culture. No detection was conducted at 5 weeks due to the lack of survived cells. RESULTS AND CONCLUSION: (1) The number of human umbilical cord blood CD34+ cells increased by MECs as the nourishing layer compared with bone marrow mesenchymal stem cells as the nourishing layer at 1, 2, and 4 weeks; however, there was no significant difference between the two groups (P > 0.05). (2) The cell immunophenotype by flow cytometry analysis indicated that only in the 2nd week, the expression of CD34+CD33- in human umbilical cord blood CD34+ cells in the control group was significantly higher in the experimental group (P 0.05). (3) The colony formation capacity of hematopoietic stem/progenitor cells showed no significant difference between the experimental and control groups at 1, 2, and 4 weeks (P > 0.05). (4) Due to the non-nourishing layer culture system, the number of human umbilical cord blood CD34+ cells decreased significantly in the 1st week, and no cells survived in the 2nd week. Therefore, blood cell immunophenotype and colony analysis could not be performed. (5) To conclude, human skeletal muscle MECs as trophoblasts are the same as human bone marrow mesenchymal stem cells, which have a hematopoietic support in vitro.

In vitro hematopoietic support of human umbilical cord blood cd34+ cells by human skeletal muscle-derived pericytes/perivascular cells / 中国组织工程研究

BACKGROUND: Perivascular cells have been shown to be the precursor cells of mesenchymal stem cells, which regulate the behavior of hematopoietic stem cells and support hematopoiesis through cell-to-cell contact or paracrine effects. Hematopoietic support of human skeletal muscle-derived pericytes/perivascular cells (hMD-PCs) remains to be studied. OBJECTIVE: To identify the biological characteristics of hMD-PCs isolated from human skeletal muscle and to study their supporting effect on umbilical cord blood CD34+ cells in vitro. METHODS: (1) hMD-PCs with phenotype CD146+ CD56-CD34-CD144-CD45- were sorted from human skeletal muscle by enzymatic digestion and multiparameter fluorescence-activated cell sorting, and their biological characteristics were identified. (2) The in vitro culture system of umbilical cord blood CD34+ cells co-cultured with human CD146+ hMD-PCs (experimental group) or with human bone marrow mesenchymal stem cells (positive control group) was established. After 1, 2 and 4 weeks of co-culture, the number of cells, the colony formation ability and immunophenotype were measured and statistically analyzed. RESULTS AND CONCLUSION: (1) CD146+ hMD-PCs were sorted by multiparameter fluorescence-activated cell sorting and the purity was (91.5±1.85)% (n=5). CD146+ hMD-PCs expressed mesenchymal surface markers CD73, CD90, CD105, CD44, and did not express hematopoietic cell and endothelial cell markers CD45, CD34, and CD31. After induced culture, CD146+ hMD-PCs could differentiate into osteoblasts, chondrogenesis, adipocytes and myoblasts. (2) There were no significant differences in the cell number, colony f ormation ability or immunophenotype (CD45+, CD34+ CD33-, CD14+, CD10+/CD19+) between experimental and positive control groups (P > 0.05, n=6). The number of cells in the blank control group without feeder was significantly decreased at 1 week of culture, and there was almost no cell survival at 2 weeks of culture. (3) In summary, CD146+ hMD-PCs, like human bone marrow mesenchymal stem cells, have hematopoietic support capacity in vitro.

Matched-sibling donor versus unrelated umbilical cord blood transplantation for treating hematological malignancies in children / 中国组织工程研究

BACKGROUND: In recent years, umbilical cord blood has gradually become a crucial alternative source of stem cells for related and unrelated bone marrow or peripheral blood hematopoietic stem cell transplantation, which is increasingly used in the treatment of hematological malignancies in children. OBJECTIVE: To compare the clinical efficacy of sibling donor umbilical cord blood transplantation and unrelated umbilical cord blood transplantation for treating hematological malignancies in children. METHODS: The clinical data of children with hematological malignancies who received umbilical cord blood transplantation at the Hematopoietic Stem Cell Transplantation Center of the First Affiliated Hospital of Zhengzhou University between January 1, 1998 and December 31, 2018 was retrospectively analyzed. All the patients received myelablative conditioning regimen, and cyslosporine A combined with or without mycophenolate mofetil were concurrently adopted for graft-versus-host disease prophylaxis. RESULTS AND CONCLUSION: (1) Two patients in the sibling donor umbilical cord blood transplantation group and three in the unrelated umbilical cord blood transplantation group did not attain hematological engraftment and subsequently died from infection, and other patients succeeded in hematological engraftment. The median time of neutrophil and platelet engraftment in the sibling donor umbilical cord blood transplantation and unrelated umbilical cord blood transplantation groups was [17 days (11-43 days), 18 days (12-45 days), P=0.307] and [20.5 days (15-50 days), 27 days (18-56 days), P=0.773]. There was no significant difference between the two groups. (2) The incidence of acute graft-versus-host disease and chronic graft-versus-host disease in the sibling donor umbilical cord blood transplantation and unrelated umbilical cord blood transplantation groups was 36% vs. 43% (P=0.737) and 15% vs. 33% (P=0.412). There was no significant difference between the two groups. There was also no significant difference in the incidence of infection after transplantation between sibling donor umbilical cord blood transplantation and unrelated umbilical cord blood transplantation groups (56% vs. 71%, P=0.343). (3) There were no significant differences in the 2-year overall survival (61% vs. 36%, P=0.301), or 2-year relapse-free survival (56% vs. 33%, P=0.151). The 5-year overall survival and 5-year relapse-free survival in the sibling donor umbilical cord blood transplantation and unrelated umbilical cord blood transplantation groups were 54% vs. 24% (P=0.044) and 50% vs. 20% (P=0.039). The results showed that there was a significant difference in long-term survival rate between two groups. (4) Our results reveal that both sibling donor umbilical cord blood transplantation and unrelated umbilical cord blood transplantation are safe, effective and applicable for children with hematological malignancies. In particular, there are significant benefits in the long-term survival of substitute donor transplantation for pediatric patients with hematological malignancies.

Effects of umbilical cord blood-derived mesenchymal stem cell transplantation on recovery of nerve function and telomerase reverse transcriptase expression in brain tissue of cerebral infarction rats / 中国组织工程研究

BACKGROUND: Umbilical cord blood-derived mesenchymal stem cells can penetrate the ependyma into the brain tissue via the systemic circulation, migrate into the injury area and differentiate into neurons. They are as substitute cells applied in the treatment of central nervous system diseases OBJECTIVE: To explore the effects of umbilical cord blood-derived mesenchymal stem cell transplantation on the functional recovery of nerve and expression of telomerase reverse transcriptase in the brain tissue of cerebral infarction rats. METHODS: One hundred and thirty rats were enrolled and were randomly divided into control group (n=30), model group (n=30), umbilical cord blood-derived mesenchymal stem cell 1 group (n=35) and umbilical cord blood-derived mesenchymal stem cell 2 group (n=35). The control group was not given any treatment. The rat models of cerebral infarction were made by internal carotid artery suture method in the other groups. At 1 and 4 days after successful modeling, 2×106 umbilical cord blood-derived mesenchymal stem cells were transplanted into tail vein in the umbilical cord blood-derived mesenchymal stem cell 1 and 2 groups. The levels of reactive oxygen species and superoxide dismutase were detected at 24 hours after transplantation. The nerve function score was detected by Y-maze test at 7 and 14 days after transplantation. The neuronal apoptosis in the center of lesions was detected by TUNEL assay. The expression levels of Caspase-3, Bax, Bcl-2 and telomerase reverse transcriptase proteins around the infarct lesion were detected by western blot assay. The mRNA expression of telomerase reverse transcriptase around infarct lesions was detected by RT-PCR. RESULTS AND CONCLUSION: Compared with the control group, in the model group, the error number in the Y-maze test, nerve function scores, apoptotic index, expression levels of Caspase-3 and Bax, and level of reactive oxygen species were significantly increased (P < 0.05), the expression of Bcl-2 and telomerase reverse transcriptase protein and mRNA, and level of superoxide dismutase were significantly decreased (P < 0.05). Compared with the model group, in the umbilical cord blood-derived mesenchymal stem cell 1 and 2 groups, the error number in the Y-maze test, nerve function score, apoptotic index, expression of Caspase-3 and Bax, and level of reactive oxygen species were significantly decreased (P < 0.05), while the expression of Bcl-2 and telomerase reverse transcriptase protein and mRNA and level of superoxide dismutase were significantly increased (P < 0.05). Compared with the umbilical cord blood-derived mesenchymal stem cell 2 group, in the umbilical cord blood-derived mesenchymal stem cell 1 group, the error number in the Y-maze test, nerve function score, apoptotic index, expression of Caspase-3 and Bax, and level of reactive oxygen species were significantly decreased (P < 0.05), while the expression of Bcl-2 and telomerase reverse transcriptase protein and mRNA and level of superoxide dismutase were significantly increased (P < 0.05). These findings indicate that umbilical cord blood-derived mesenchymal stem cell transplantation can effectively promote the neurological recovery in rats with cerebral infarction, and up-regulate the expression of telomerase reverse transcriptase in rat brain tissue. Moreover, earlier cell transplantation indicates better effects.

Transplantation of human umbilical cord blood mesenchymal stem cells in the repair of hematopoietic injury in bone marrow / 中国组织工程研究

BACKGROUND: A large number of studies mainly concern the proliferation effect of mesenchymal stem cells on hematopoietic stem cells in vitro and that bone marrow mesenchymal stem cell transplantation can reduce the death of hematopoietic cells caused by irradiation, increase the survival of bone marrow cells and repair hematopoiesis, while few of them investigate the repair of human umbilical cord blood mesenchymal stem cells transplantation on bone marrow hematopoiesis injury. OBJECTIVE: To explore the repair of hematopoietic microenvironment of bone marrow by human umbilical cord blood mesenchymal stem cells. METHODS: Male BALB/c mice were randomly divided into three groups. The mice in experimental group and control group were irradiated with total dose of 6-Gy X-ray to establish a mouse model of bone marrow hematopoietic injury. The normal group contained untreated normal mice. In the experimental group, CM-DiL labeled human umbilical cord blood mesenchymal stem cells were injected into the tail vein of each mouse at 5×106 (0.2 mL). The control group and the normal group received normal saline 0.2 mL through the tail vein. The peripheral blood hematology and bone marrow hematopoietic microenvironment repair were observed at 1, 5, 7, 14 and 21 days after cell transplantation. RESULTS AND CONCLUSION: Peripheral blood condition: At 1, 5 and 7 days after transplantation, the leucocyte, platelet, erythrocyte count and hemoglobin concentration in the experimental group and control group decreased progressively compared with the normal group. The most obvious decrease occurred on day 7. The trilineage recovered on day 14 after transplantation, basically returned to normal on day 21 after transplantation. Compared with the experimental group, the decrease of the trilineage in the control group was more obvious. The recovery was obvious faster in the experimental group than in the control group on day 14 after transplantation. Bone marrow smears: Bone marrow smears showed that the hematopoietic function was inhibited in the experimental group and the control group at 1, 5, 7, and 14 days after transplantation, especially on day 7. Bone marrow proliferation recovered on day 14 after transplantation. It was better in the experimental group than in the control group. On day 21 after transplantation, the hematopoietic function of bone marrow of mice in the experimental group and the control group recovered, and there was no difference between the experimental group and the control group compared with the normal group. Bone marrow pathological section: Bone marrow pathological sections showed that at 1, 5, 7, and 14 days after transplantation, the hematopoietic function of bone marrow in the experimental group and the control group was inhibited. On day 14 after transplantation, the bone marrow hematopoietic function of the experimental group and the control group began to recover, but the bone marrow proliferation of the experimental group was better than that of the control group. On day 21 after transplantation, there was no difference in the bone marrow proliferation between the experimental and the control groups and the normal group. The results suggested that human umbilical cord blood mesenchymal stem cells can promote the recovery of hematopoietic function of bone marrow.