BACKGROUND: Exogenous neural stem cell transplantation promotes neural regeneration. However, various types of stem cells transplantation outcomes remain controversial. OBJECTIVE: To explore distribution, proliferatio...BACKGROUND: Exogenous neural stem cell transplantation promotes neural regeneration. However, various types of stem cells transplantation outcomes remain controversial. OBJECTIVE: To explore distribution, proliferation and differentiation of human neural stem cells (hNSCs) and human umbilical cord blood stem cells (hUCBSCs) following transplantation in ischemic brain tissue of rats, and to compare therapeutic outcomes between hNSCs and hUCBSCs. DESIGN, TIME AND SETTING: Randomized controlled animal studies were performed at the Experimental Animal Center of Nanjing Medical University and Central Laboratory of Second Affiliated Hospital of Nanjing Medical University of China from September 2008 to April 2009. MATERIALS: hNSCs were harvested from brain tissue of 10-13 week old fetuses following spontaneous abortion, and hUCBSCs were collected from umbilical cord blood of full-term newborns at the Second Affiliated Hospital of Nanjing Medical University of China. hNSCs and hUCBSCs were labeled by 5-bromodeoxyuridine (BrdU) prior to transplantation. METHODS: Rat models of cerebral ischemia were established by the suture method. A total of 60 healthy male Sprague Dawley rats aged 7-9 weeks were randomly assigned to hNSC transplantation, hUCBSC transplantation and control groups. The rat models in the hNSC transplantation, hUCBSC transplantation and control groups were infused with hNSC suspension, hUCBSC suspension and saline via the caudal vein, respectively. MAIN OUTCOME MEASURES: The distribution, proliferation and differentiation of hNSCs and hUCBSCs in ischemic brain tissue were observed using immunohistochemical methods. Neurological function in rats was assessed using the neurological severity score. RESULTS: The number of BrdU-positive cells was significantly greater in the hNSC transplantation group compared with hUCBSC transplantation group at 14 days following transplantation (P < 0.05). The number of BrdU-positive cells reached a peak at 28 days following transplantation. Nestin-positive, glial fibrillary acidic protein-positive, cyclic nucleotide 3' phosphohydrolase-positive and neuron specific enolase-positive cells were visible following transplantation. No significant difference was determined in the constituent ratio of various cells between hNSC and hUCBSC transplantation groups (P > 0.05). The neurological severity score was significantly decreased in rats at 21 days following transplantation (P < 0.05). No significant difference was detected in neurological severity score between hNSC and hUCBSC transplantation groups at various time points (P > 0.05). CONCLUSION: The transplanted hNSCs and hUCBSCs can migrate into ischemic brain tissue, proliferate and differentiate into neuron-like, astrocyte-like and oligodendrocyte-like cells, and improve neurological function in rats with cerebral ischemia.展开更多
BACKGROUND: Transplanted mononuclear cell(MNC)of umbilical blood can survive in central nervous system (CNS)of host through blood brain barrier,differentiate into nerver cells,migrate to damaged site and integrate mor...BACKGROUND: Transplanted mononuclear cell(MNC)of umbilical blood can survive in central nervous system (CNS)of host through blood brain barrier,differentiate into nerver cells,migrate to damaged site and integrate morphological strucgh and function with nerve cells of host so as to improve deficiencies of sensatory function,motor function and cognitive function and influence on stroke sequela.OBJECTIVE: To observe the vein transplantation of human umbilical cord blood stem cells(HUCBSC) for improving neurological function,limb funtion and activity of daily living of patients with stroke and evaluate the reliability.DESIGN: Self-controlled study.SETTING: Department of Neurosurgery,the Second People's Hospital of Zhengzhou City;Red-crossed Blood Center of Henan Province;Department of Neurosurgery,the Fist Affiliated Hospital of Zhenzhou University.PARTICIPANTS:A total of 10 patients with stoke sequela were selected from Department of Cerebral Surgery,the Second People's Hospital of Zhengzhou City from April to December 2005.There were 9males and 1 female aged from 35to 75years with the mean age of 56 years.All of them were diagnosed with CT and MRI examination and coincidence with diagnostic criteria of stroke established by the Fourth National Academic Meeting for Cerebrovascular Disease.All patients provided informed consent. METHODS:80-140 mL umbilical blood of term birth of newborn was selected hermetically and maintained in sterile plastic bag.And then,the blood was centrifugated at the speed of 1500 r/min for 30 minutes at 22℃ in order to separate MNC,i.e.,HUCBSC.In addition,after final diagnosis during hospitalization,stroke patients were perfused with HUCBSC through superficial vein of back of the hand.Each patient was averagely penfused with 6 portions of HUCBSC(cellular numbers≥1×108/portion)and the interval between each portion was 1-7 days with the mean interval of 4 days.MAIN OUTCOME MEASURES: ①Neurological function of stroke patients was evaluated with neurological function deficiency(NFD)before treatment and at 3 months after treatment.The scale includes consciousness,level fix function.facial paralysis,language,muscle force of upper limbs,muscle force of lower limb and step function.The total scores ranged from 0 to 45; meanwhile,the lower the scores were,the better the neurological function of limbs was. ②Motor function of injured limbs was evaluated with Fugl-Meyer Assessment (FMA),including motor function of upper limbs.motor function of lower limbs,balance ability,sensory function and motion of joint.The total scores ranged from 0 to 226;meanwhile,the higher the scores were,the better the motor function of limbs was.③Activities of daily living(ADL)was evaluated with Barthel Index(BI),including having meals,taking a bath,dressing oneself,putting on clothes,walking in balance and stair activity.The total scores ranged from 0 to 100;meanwhile,the higher the scores were,the stronger the ADL was.RESULTS:A total of 10 patients were involved in the final analysis.After treatment,NFD of stroke patients was (10.9±5.09)points,which was lower than that before treatment[(25.4±6.09)points,t=8.213,P＜0.01].In addition,after treatment,FMA and BI of stroke patients were(80.9±25.00)points and(81.1±15.93)points,re spectively,which were higher than those before treatment[(31.9+21.85)points,(36.2+19.41)points,t=13.024,13.670,P＜0.01]. Immuno-suppressive drugs were not used during the whole therapeutic procedure;moreover,immunological rejection and allergic reaction were not observed during the same period.CONCLUSION:Transplanting HUCBSC through superficial vein of back of the hand is regarded as a simple and safe method for the treatment of stroke sequela.展开更多
Umbilical cord blood(UCB)is a primitive and abundant source of mesenchymal stem cells(MSCs).UCB-derived MSCs have a broad and efficient therapeutic capacity to treat various diseases and disorders.Despite the high lat...Umbilical cord blood(UCB)is a primitive and abundant source of mesenchymal stem cells(MSCs).UCB-derived MSCs have a broad and efficient therapeutic capacity to treat various diseases and disorders.Despite the high latent selfrenewal and differentiation capacity of these cells,the safety,efficacy,and yield of MSCs expanded for ex vivo clinical applications remains a concern.However,immunomodulatory effects have emerged in various disease models,exhibiting specific mechanisms of action,such as cell migration and homing,angiogenesis,anti-apoptosis,proliferation,anti-cancer,anti-fibrosis,anti-inflammation and tissue regeneration.Herein,we review the current literature pertaining to the UCB-derived MSC application as potential treatment strategies,and discuss the concerns regarding the safety and mass production issues in future applications.展开更多
Clinical and laboratory results document psoriatic arthritis in a 56-year old patient. The symptoms did not resolve with standard treatments(nonsteroidal anti-inflammatory drugs, steroids and methotrexate). TNF-alpha ...Clinical and laboratory results document psoriatic arthritis in a 56-year old patient. The symptoms did not resolve with standard treatments(nonsteroidal anti-inflammatory drugs, steroids and methotrexate). TNF-alpha inhibitors(certolizumab pegol and adalimumab) were added to the treatment regime, with some adverse effects. A trial of human umbilical cord stem cell therapy was then initiated. The stem cells were enriched and concentrated from whole cord blood, by removal of erythrocytes and centrifugation. The patient received several infusions of cord blood stem cells, through intravenous and intra-articular injections. These stem cell treatments correlated with remission of symptoms(joint pain and psoriatic plaques) and normalized serologic results for the inflammatory markers C-reactive protein and erythrocyte sedimentation rate. These improvements were noted within the first thirty days post-treatment, and were sustained for more than one year. The results of this trial suggest that cord blood stem cells may have important therapeutic value for patients with psoriatic arthritis, particularly for those who cannot tolerate standard treatments.展开更多
Spinal cord injury(SCI)continues to be a pressing health and social problem.The injury leads to neuronal and glial cell death accompanied by degeneration of nerve fibers.There are currently no particularly effective t...Spinal cord injury(SCI)continues to be a pressing health and social problem.The injury leads to neuronal and glial cell death accompanied by degeneration of nerve fibers.There are currently no particularly effective treatments.SCI causes profound disability of people affected and has attracted increased attention in the international field of neuroregeneration.For the past two decades,展开更多
BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human gra...BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1(2×108 L-1,5×108 L-1,1×109 L-1)], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=(1-A experimental well-A effector cell well/A target cell well)×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was (43.12±5.83)%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[(31.00 ±4.41)%,(30.92±5.27)%,(33.57±5.35)%,(26.23±5.20)%, t=3.51,2.98,4.24, P < 0.01); But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 (t=2.01,2.36, P < 0.05), and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1(t=0.06,P > 0.05). CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.展开更多
Objective: In this study, we aimed to investigate umbilical cord blood CD33 and erythropoietin (EPO) levels of pregnants with abnormal umbilical and uterine artery doppler waveforms and to compare with normal pregnanc...Objective: In this study, we aimed to investigate umbilical cord blood CD33 and erythropoietin (EPO) levels of pregnants with abnormal umbilical and uterine artery doppler waveforms and to compare with normal pregnancies. Materials and Methods: Total 40 pregnant women were included in this study. Of these 40 women, while 20 patients had abnormal umbilical and uterine artery doppler waveforms, the other 20 patients had normal umbilical and uterine artery doppler waveforms. After the delivery, blood samples were taken from umbilical artery of double clemped umbilical cord for blood gas parameters, EPO and CD33 levels. Sociodemographic findings, antepartum, intrapartum test results, labor and delivery characteristics and newborn examination results were recorded. Blood gas parameters, EPO and CD33 levels between groups were analyzed. Mann-Whitney U test and t-test were used as statistical methods. Results: There were no differences between parity, gestational ages and newborn weights of the groups. Cord blood CD33 and EPO levels of group with abnormal umbilical and uterine artery doppler waveforms were significantly higher than group with normal umbilical and uterine artery doppler waveforms (p < 0.01). Conclusion: Pathology on doppler screen shows to us a connection between chronic hypoxemia and abnormal on doppler screen. Preference of high blood CD33 levels for cord blood transplantation especially during last years can also be used with preference of cord blood with abnormal doppler findings.展开更多
Umbilical cord blood(UCB)is an efficient and valuable source of hematopoietic stem cells(HSCs)for transplantation.In addition to HSCs it harbours low amounts of mesenchymal stem cells(MSCs).No single marker to identif...Umbilical cord blood(UCB)is an efficient and valuable source of hematopoietic stem cells(HSCs)for transplantation.In addition to HSCs it harbours low amounts of mesenchymal stem cells(MSCs).No single marker to identify cord blood-derived stem cells,or to indicate their multipotent phenotype,has been characterized so far.SSEA-3 and-4 are cell surface globoseries glycosphingolipid epitopes that are commonly used as markers for human embryonic stem cells,where SSEA-3 rapidly disappears when the cells start to differentiate.Lately SSEA-3 and-4 have also been observed in MSCs.As there is an ongoing discussion and variation of stem-cell markers between laboratories,we have now comprehensively characterized the expression of these epitopes in both the multipotent stem-cell types derived from UCB.We have performed complementary analysis using gene expression analysis,mass spectrometry and immunochemical methods,including both flow cytometry and immunofluoresence microscopy.SSEA-4,but not SSEA-3,was expressed on MSCs but absent from HSCs.Our findings indicate that SSEA-3 and/or-4 may not be optimal markers for multipotency in the case of stem cells derived from cord blood,as their expression may be altered by cell-culture conditions.展开更多
The effects of hematopoietic stem/progenitor cells(HSPCs)expanded in the two step coculture with human bone marrow mesenchymal stem cells(hMSCs)on the hematopoietic reconstruction of irradiated NOD/SCID mice were stud...The effects of hematopoietic stem/progenitor cells(HSPCs)expanded in the two step coculture with human bone marrow mesenchymal stem cells(hMSCs)on the hematopoietic reconstruction of irradiated NOD/SCID mice were studied.Mononuclear cells(MNCs)were isolated from human umbilical cord blood(UCB)and cultured in the non-coculture scheme of rhSCF+rhG−CSF+rhMDGF combination and the coculture scheme of rhSCF+rhG−CSF+rhMDGF+hMSCs.Sublethally-irradiated NOD/SCID mice were transplanted with ex vivo expanded HSPCs with the dose of 8.5×10^(6) cells per mouse.After transplantation,the dynamics of WBC in the transplanted mice was measured periodically,and the Alu sequence fragment special for human in the transplanted mice was inspected by PCR.Results showed that the coculture scheme increased proliferation of UCB-derived HSPCs.After transplantation with expanded HSPCs,the population of WBC in the transplanted mice increased in 12 d and reached the first peak in 25 d,then showed the second increasing of WBC in 45~55 d.Expanded cells from the coculture scheme appeared to be favorable for the second increasing of WBC in the transplanted mice.After 85 d,the Alu sequence fragment was detected in the probability of 87.5%(7/8)for the non-coculture scheme and 88.9%(8/9)for the coculture scheme.展开更多
Objective:To evaluate the predictive ability of neonate condition through the traditional parameters and artery umbilical cord blood gas(aUCBG).Methods:A prospective cohort study was conducted in obstetrics and gyneco...Objective:To evaluate the predictive ability of neonate condition through the traditional parameters and artery umbilical cord blood gas(aUCBG).Methods:A prospective cohort study was conducted in obstetrics and gynecology department between October 2017 and August 2018 at Tongji Hospital in Wuhan,China,and 360 aUCBG samples were collected.The average age of pregnant women was(29.50±4.42)years,range from 19 to 48 years old.The gestational age range from 28+4 weeks to 41+3 weeks at admission.Logistic regression and area under the curve(AUC)from Receiver operating characteristic curves were used to identify risk factors,such as,premature rupture of membranes(PROM),high blood pressure,premature delivery(PD),low 1-minute Apgar scores(Apgar 1),low 5-minute Apgar scores(Apgar 5),pH,base excess,bicarbonate,neonatal blood sugar(NBS),and so on,to predict neonatal condition and evaluate the predictive ability of traditional and aUCBG parameters.Results:In all cases,PROM,PD,Apgar 1,Apgar 5,pH,base excess,bicarbonate,total carbon dioxide,and neonatal blood sugar were risk factors and were associated with poor condition of neonate.Apgar 1 were an independent risk factor.Combined traditional and aUCBG parameters had higher AUC of 0.895(95%confidence interval(C/):0.830-0.960,P<0.001).In cesarean section subgroup,high blood pressure,PD,and Apgar 1 were risk factors and were associated with poor condition of neonate.Apgar 1 and low pH were the independent risk factors.Combined traditional and aUCBG parameters had highest AUC of 0.940(95%C/:0.886-0.993,P<0.001).In vaginal delivery subgroup,maternal age above 35 years,PROM,PD,Apgar 1,Apgar 5,and male newborn were risk factors and were associated with poor condition of neonate.Maternal age above 35 years was an independent risk factor.Combined traditional and aUCBG parameters had highest AUC of 0.897(95%Cl:0.828-0.965,P<0.001).For pregnant women without comorbidities and complications of pregnancy,aUCBG may not be necessat7.Conclusion:In high-risk pregnancies,especially lower Apgar scores,PD,and maternal age above 35-year old,aUCBG is recommended.Traditional parameters combined with aUCBG might increase the predicting ability of neonate condition.展开更多
OBJECTIVE: The aim of this study was to evaluate the effectiveness and safety of stem cell transplantation for spinal cord injury(SCI).DATA SOURCES: PubM ed, EMBASE, Cochrane, China National Knowledge Infrastructure, ...OBJECTIVE: The aim of this study was to evaluate the effectiveness and safety of stem cell transplantation for spinal cord injury(SCI).DATA SOURCES: PubM ed, EMBASE, Cochrane, China National Knowledge Infrastructure, China Science and Technology Journal, Wanfang, and Sino Med databases were systematically searched by computer to select clinical randomized controlled trials using stem cell transplantation to treat SCI, published between each database initiation and July 2016. DATA SELECTION: Randomized controlled trials comparing stem cell transplantation with rehabilitation treatment for patients with SCI. Inclusion criteria:(1) Patients with SCI diagnosed according to the American Spinal Injury Association(ASIA) International standards for neurological classification of SCI;(2) patients with SCI who received only stem cell transplantation therapy or stem cell transplantation combined with rehabilitation therapy;(3) one or more of the following outcomes reported: outcomes concerning neurological function including sensory function and locomotor function, activities of daily living, urination functions, and severity of SCI or adverse effects. Studies comprising patients with complications, without full-text, and preclinical animal models were excluded. Quality of the included studies was evaluated using the Cochrane risk of bias assessment tool and Rev Man V5.3 software, provided by the Cochrane Collaboration, was used to perform statistical analysis. OUTCOME MEASURES: ASIA motor score, ASIA light touch score, ASIA pinprick score, ASIA impairment scale grading improvement rate, activities of daily living score, residual urine volume, and adverse events.RESULTS: Ten studies comprising 377 patients were included in the analysis and the overall risk of bias was relatively low level. Four studies did not detail how random sequences were generated, two studies did not clearly state the blinding outcome assessment, two studies lacked blinding outcome assessment, one study lacked follow-up information, and four studies carried out selective reporting. Compared with rehabilitation therapy, stem cell transplantation significantly increased the lower limb light touch score(odds ratio(OR) = 3.43, 95% confidence interval(CI): 0.01 – 6.86, P = 0.05), lower limb pinprick score(OR = 3.93, 95%CI: 0.74 – 7.12, P = 0.02), ASI grading rate(relative risk(RR) = 2.95, 95%CI: 1.64 – 5.29, P = 0.0003), and notably reduced residual urine volume(OR = –8.10, 95%CI: –15.09 to –1.10, P = 0.02). However, stem cell transplantation did not significantly improve motor score(OR = 1.89, 95%CI: –0.25 to 4.03, P = 0.08) or activities of daily living score(OR = 1.12, 95%CI: –1.17 to 4.04, P = 0.45). Furthermore, stem cell transplantation caused a high rate of mild adverse effects(RR = 14.49, 95%CI: 5.34 – 34.08, P < 0.00001); however, these were alleviated in a short time. CONCLUSION: Stem cell transplantation was determined to be an efficient and safe treatment for SCI and simultaneously improved sensory and bladder functions. Although associated minor and temporary adverse effects were observed with transplanted stem cells, spinal cord repair and axon remyelination were apparent. More randomized controlled trials with larger sample sizes and longer follow-up times are needed to further validate the effectiveness of stem cell transplantation in the treatment of SCI.展开更多
Despite emerging contemporary biotechnological methods such as gene-and stem cell-based therapy,there are no clinically established therapeutic strategies for neural regeneration after spinal cord injury.Our previous ...Despite emerging contemporary biotechnological methods such as gene-and stem cell-based therapy,there are no clinically established therapeutic strategies for neural regeneration after spinal cord injury.Our previous studies have demonstrated that transplantation of genetically engineered human umbilical cord blood mononuclear cells producing three recombinant therapeutic molecules,including vascular endothelial growth factor(VEGF),glial cell-line derived neurotrophic factor(GDNF),and neural cell adhesion molecule(NCAM)can improve morpho-functional recovery of injured spinal cord in rats and mini-pigs.To investigate the efficacy of human umbilical cord blood mononuclear cells-mediated triple-gene therapy combined with epidural electrical stimulation in the treatment of spinal cord injury,in this study,rats with moderate spinal cord contusion injury were intrathecally infused with human umbilical cord blood mononuclear cells expressing recombinant genes VEGF165,GDNF,NCAM1 at 4 hours after spinal cord injury.Three days after injury,epidural stimulations were given simultaneously above the lesion site at C5(to stimulate the cervical network related to forelimb functions)and below the lesion site at L2(to activate the central pattern generators)every other day for 4 weeks.Rats subjected to the combined treatment showed a limited functional improvement of the knee joint,high preservation of muscle fiber area in tibialis anterior muscle and increased H/M ratio in gastrocnemius muscle 30 days after spinal cord injury.However,beneficial cellular outcomes such as reduced apoptosis and increased sparing of the gray and white matters,and enhanced expression of heat shock and synaptic proteins were found in rats with spinal cord injury subjected to the combined epidural electrical stimulation with gene therapy.This study presents the first proof of principle study of combination of the multisite epidural electrical stimulation with ex vivo triple gene therapy(VEGF,GDNF and NCAM)for treatment of spinal cord injury in rat models.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.2.20.02.18)on February 20,2018.展开更多
BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferati...BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferating potential in kidney injury in mice.METHODS Human umbilical cord blood(UCB)-derived CD34+cells were incubated for one week in vasculogenic conditioning medium.Vasculogenic culture significantly increased the number of CD34+cells and their ability to form endothelial progenitor cell colony-forming units.Adenineinduced tubulointerstitial injury of the kidney was induced in immunodeficient non-obese diabetic/severe combined immunodeficiency mice,and cultured human UCB-CD34+cells were administered at a dose of 1×106/mouse on days 7,14,and 21 after the start of adenine diet.RESULTS Repetitive administration of cultured UCB-CD34+cells significantly improved the time-course of kidney dysfunction in the cell therapy group compared with that in the control group.Both interstitial fibrosis and tubular damage were significantly reduced in the cell therapy group compared with those in the control group(P<0.01).Microvasculature integrity was significantly preserved(P<0.01)and macrophage infiltration into kidney tissue was dramatically decreased in the cell therapy group compared with those in the control group(P<0.001).CONCLUSION Early intervention using human cultured CD34+cells significantly improved the progression of tubulointerstitial kidney injury.Repetitive administration of cultured human UCB-CD34+cells significantly improved tubulointerstitial damage in adenine-induced kidney injury in mice via vasculoprotective and anti-inflammatory effects.展开更多
Tay-Sachs disease and Sandhoff disease are severe hereditary neurodegenerative disorders caused by a deficiency ofβ-hexosaminidase A(HexA)enzyme,which results in the accumulation of GM2 gangliosides in the nervous sy...Tay-Sachs disease and Sandhoff disease are severe hereditary neurodegenerative disorders caused by a deficiency ofβ-hexosaminidase A(HexA)enzyme,which results in the accumulation of GM2 gangliosides in the nervous system cells.In this work,we analyzed the efficacy and safety of cell-mediated gene therapy for Sandhoff disease and Sandhoff disease using a bicistronic lentiviral vector encoding cDNA of HexAα-andβ-subunit genes separated by the nucleotide sequence of a P2A peptide(HEXA-HEXB).The functionality of the bicistronic construct containing the HEXA-HEXB genetic cassette was analyzed in a culture of HEK293T cells and human umbilical cord blood mononuclear cells(hUCBMCs).Our results showed that the enzymatic activity of HexA in the conditioned medium harvested from genetically modified HEK293T-HEXA-HEXB and hUCBMCs-HEXA-HEXB was increased by 23 and 8 times,respectively,compared with the conditioned medium of native cells.Western blot analysis showed that hUCBMCs-HEXA-HEXB secreted both completely separated HEXA and HEXB proteins,and an uncleaved protein containing HEXA+HEXB linked by the P2A peptide.Intravenous injection of genetically modified hUCBMCs-HEXA-HEXB to laboratory Wistar rats was carried out,and the HexA enzymatic activity in the blood plasma of experimental animals,as well as the number of live cells of immune system organs(spleen,thymus,bone marrow,lymph nodes)were determined.A significant increase in the enzymatic activity of HexA in the blood plasma of laboratory rats on days 6 and 9(by 2.5 and 3 times,respectively)after the administration of hUCBMCsHEXA-HEXB was shown.At the same time,the number of live cells in the studied organs remained unchanged.Thus,the functionality of the bicistronic genetic construct encoding cDNA of the HEXA and HEXB genes separated by the nucleotide sequence of the P2A peptide was shown in vitro and in vivo.We hypothesize that due to the natural ability of hUCBMCs to overcome biological barriers,such a strategy can restore the activity of the missing enzyme in the central nervous system of patients with GM2 gangliosidoses.Based on the obtained data,it can be concluded that intravenous administration of hUCBMCs with HexA overexpression is a promising method of the therapy for GM2 gangliosidoses.The animal protocol was approved by the Animal Ethics Committee of the Kazan Federal University(No.23)on June 30,2020.展开更多
Autism spectrum disorder(ASD)is a complex neurodevelopmental condition characterized by impairments in social communication,abnormal to absent verbal communication,the presence of repetitive stereotypic verbal and non...Autism spectrum disorder(ASD)is a complex neurodevelopmental condition characterized by impairments in social communication,abnormal to absent verbal communication,the presence of repetitive stereotypic verbal and nonverbal behaviors and restricted interests,with onset early in life.We showed cognitive and behavioral characteristics of ASD by impairment of communication,cognition,perception,motor skills,executive function,theory of mind and emotion control.Recently,pathogenesis of immune pathology in the brains of individuals with ASD has been focused.New therapeutic approaches in the viewpoints of immune modulation and microglial function are logical for novel treatments for individuals with ASD.Cell therapies such as umbilical cord blood cells and mesenchymal stromal cells for ASD will be a challenging and encouraging field in the future clinical study with progress of biotechnological science.展开更多
Diabetic patients often exhibit delayed or incomplete progress in the healing of acute wounds,owing to poor blood perfusion.Platelet-rich plasma(PRP)has attracted much attention as a means to improve wound healing,bec...Diabetic patients often exhibit delayed or incomplete progress in the healing of acute wounds,owing to poor blood perfusion.Platelet-rich plasma(PRP)has attracted much attention as a means to improve wound healing,because it contains high growth factor concentrations.However,the burst-like release of PRP growth factors results in a short halflife of these therapeutic proteins,thus greatly limiting the therapeutic effect.In this study,we prepared PRP from human umbilical cord blood and developed an in situ photocrosslinkable PRP hydrogel glue(HNPRP)by adding a photoresponsive hyaluronic acid(HA-NB)into PRP.The HNPRP hydrogel allowed for controlled release of plateletderived growth factor-BB(PDGF-BB)and transforming growth factor-β(TGF-β)for up to 28 days.In vitro cell culture showed that HNPRP promoted migration of fibroblasts and keratinocytes as well as PRP and did not reveal the advantages of HNPRP.However,in a diabetic rat skin wound model,HNPRP treatment promoted faster wound closure.Furthermore,the HNPRP group,compared with the control,PRP and hydrogel only groups,showed significantly greater re-epithelialization and numbers of both newly formed and mature blood vessels.The HNPRP group also displayed higher collagen formation than did the control group.In conclusion,HNPRP enhances angiogenesis and skin regeneration and consequently achieves faster wound healing,thus extending its potential for clinical applications to treat diabetic skin wounds.展开更多
文摘BACKGROUND: Exogenous neural stem cell transplantation promotes neural regeneration. However, various types of stem cells transplantation outcomes remain controversial. OBJECTIVE: To explore distribution, proliferation and differentiation of human neural stem cells (hNSCs) and human umbilical cord blood stem cells (hUCBSCs) following transplantation in ischemic brain tissue of rats, and to compare therapeutic outcomes between hNSCs and hUCBSCs. DESIGN, TIME AND SETTING: Randomized controlled animal studies were performed at the Experimental Animal Center of Nanjing Medical University and Central Laboratory of Second Affiliated Hospital of Nanjing Medical University of China from September 2008 to April 2009. MATERIALS: hNSCs were harvested from brain tissue of 10-13 week old fetuses following spontaneous abortion, and hUCBSCs were collected from umbilical cord blood of full-term newborns at the Second Affiliated Hospital of Nanjing Medical University of China. hNSCs and hUCBSCs were labeled by 5-bromodeoxyuridine (BrdU) prior to transplantation. METHODS: Rat models of cerebral ischemia were established by the suture method. A total of 60 healthy male Sprague Dawley rats aged 7-9 weeks were randomly assigned to hNSC transplantation, hUCBSC transplantation and control groups. The rat models in the hNSC transplantation, hUCBSC transplantation and control groups were infused with hNSC suspension, hUCBSC suspension and saline via the caudal vein, respectively. MAIN OUTCOME MEASURES: The distribution, proliferation and differentiation of hNSCs and hUCBSCs in ischemic brain tissue were observed using immunohistochemical methods. Neurological function in rats was assessed using the neurological severity score. RESULTS: The number of BrdU-positive cells was significantly greater in the hNSC transplantation group compared with hUCBSC transplantation group at 14 days following transplantation (P < 0.05). The number of BrdU-positive cells reached a peak at 28 days following transplantation. Nestin-positive, glial fibrillary acidic protein-positive, cyclic nucleotide 3' phosphohydrolase-positive and neuron specific enolase-positive cells were visible following transplantation. No significant difference was determined in the constituent ratio of various cells between hNSC and hUCBSC transplantation groups (P > 0.05). The neurological severity score was significantly decreased in rats at 21 days following transplantation (P < 0.05). No significant difference was detected in neurological severity score between hNSC and hUCBSC transplantation groups at various time points (P > 0.05). CONCLUSION: The transplanted hNSCs and hUCBSCs can migrate into ischemic brain tissue, proliferate and differentiate into neuron-like, astrocyte-like and oligodendrocyte-like cells, and improve neurological function in rats with cerebral ischemia.
文摘BACKGROUND: Transplanted mononuclear cell(MNC)of umbilical blood can survive in central nervous system (CNS)of host through blood brain barrier,differentiate into nerver cells,migrate to damaged site and integrate morphological strucgh and function with nerve cells of host so as to improve deficiencies of sensatory function,motor function and cognitive function and influence on stroke sequela.OBJECTIVE: To observe the vein transplantation of human umbilical cord blood stem cells(HUCBSC) for improving neurological function,limb funtion and activity of daily living of patients with stroke and evaluate the reliability.DESIGN: Self-controlled study.SETTING: Department of Neurosurgery,the Second People's Hospital of Zhengzhou City;Red-crossed Blood Center of Henan Province;Department of Neurosurgery,the Fist Affiliated Hospital of Zhenzhou University.PARTICIPANTS:A total of 10 patients with stoke sequela were selected from Department of Cerebral Surgery,the Second People's Hospital of Zhengzhou City from April to December 2005.There were 9males and 1 female aged from 35to 75years with the mean age of 56 years.All of them were diagnosed with CT and MRI examination and coincidence with diagnostic criteria of stroke established by the Fourth National Academic Meeting for Cerebrovascular Disease.All patients provided informed consent. METHODS:80-140 mL umbilical blood of term birth of newborn was selected hermetically and maintained in sterile plastic bag.And then,the blood was centrifugated at the speed of 1500 r/min for 30 minutes at 22℃ in order to separate MNC,i.e.,HUCBSC.In addition,after final diagnosis during hospitalization,stroke patients were perfused with HUCBSC through superficial vein of back of the hand.Each patient was averagely penfused with 6 portions of HUCBSC(cellular numbers≥1×108/portion)and the interval between each portion was 1-7 days with the mean interval of 4 days.MAIN OUTCOME MEASURES: ①Neurological function of stroke patients was evaluated with neurological function deficiency(NFD)before treatment and at 3 months after treatment.The scale includes consciousness,level fix function.facial paralysis,language,muscle force of upper limbs,muscle force of lower limb and step function.The total scores ranged from 0 to 45; meanwhile,the lower the scores were,the better the neurological function of limbs was. ②Motor function of injured limbs was evaluated with Fugl-Meyer Assessment (FMA),including motor function of upper limbs.motor function of lower limbs,balance ability,sensory function and motion of joint.The total scores ranged from 0 to 226;meanwhile,the higher the scores were,the better the motor function of limbs was.③Activities of daily living(ADL)was evaluated with Barthel Index(BI),including having meals,taking a bath,dressing oneself,putting on clothes,walking in balance and stair activity.The total scores ranged from 0 to 100;meanwhile,the higher the scores were,the stronger the ADL was.RESULTS:A total of 10 patients were involved in the final analysis.After treatment,NFD of stroke patients was (10.9±5.09)points,which was lower than that before treatment[(25.4±6.09)points,t=8.213,P＜0.01].In addition,after treatment,FMA and BI of stroke patients were(80.9±25.00)points and(81.1±15.93)points,re spectively,which were higher than those before treatment[(31.9+21.85)points,(36.2+19.41)points,t=13.024,13.670,P＜0.01]. Immuno-suppressive drugs were not used during the whole therapeutic procedure;moreover,immunological rejection and allergic reaction were not observed during the same period.CONCLUSION:Transplanting HUCBSC through superficial vein of back of the hand is regarded as a simple and safe method for the treatment of stroke sequela.
文摘Umbilical cord blood(UCB)is a primitive and abundant source of mesenchymal stem cells(MSCs).UCB-derived MSCs have a broad and efficient therapeutic capacity to treat various diseases and disorders.Despite the high latent selfrenewal and differentiation capacity of these cells,the safety,efficacy,and yield of MSCs expanded for ex vivo clinical applications remains a concern.However,immunomodulatory effects have emerged in various disease models,exhibiting specific mechanisms of action,such as cell migration and homing,angiogenesis,anti-apoptosis,proliferation,anti-cancer,anti-fibrosis,anti-inflammation and tissue regeneration.Herein,we review the current literature pertaining to the UCB-derived MSC application as potential treatment strategies,and discuss the concerns regarding the safety and mass production issues in future applications.
文摘Clinical and laboratory results document psoriatic arthritis in a 56-year old patient. The symptoms did not resolve with standard treatments(nonsteroidal anti-inflammatory drugs, steroids and methotrexate). TNF-alpha inhibitors(certolizumab pegol and adalimumab) were added to the treatment regime, with some adverse effects. A trial of human umbilical cord stem cell therapy was then initiated. The stem cells were enriched and concentrated from whole cord blood, by removal of erythrocytes and centrifugation. The patient received several infusions of cord blood stem cells, through intravenous and intra-articular injections. These stem cell treatments correlated with remission of symptoms(joint pain and psoriatic plaques) and normalized serologic results for the inflammatory markers C-reactive protein and erythrocyte sedimentation rate. These improvements were noted within the first thirty days post-treatment, and were sustained for more than one year. The results of this trial suggest that cord blood stem cells may have important therapeutic value for patients with psoriatic arthritis, particularly for those who cannot tolerate standard treatments.
基金supported by grants 15-04-07527(AAR) and 16-34-60101(YOM) from Russian Foundation for Basic Researchsupported by a Presidential Grant for government support of young scientists(PhD) from the Russian Federation(MK-4020.2015.7)+1 种基金performed in accordance with Program of Competitive Growth of Kazan Federal Universitya subsidy allocated to Kazan Federal University for the state assignment in the sphere of scientific activities
文摘Spinal cord injury(SCI)continues to be a pressing health and social problem.The injury leads to neuronal and glial cell death accompanied by degeneration of nerve fibers.There are currently no particularly effective treatments.SCI causes profound disability of people affected and has attracted increased attention in the international field of neuroregeneration.For the past two decades,
文摘BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1(2×108 L-1,5×108 L-1,1×109 L-1)], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=(1-A experimental well-A effector cell well/A target cell well)×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was (43.12±5.83)%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[(31.00 ±4.41)%,(30.92±5.27)%,(33.57±5.35)%,(26.23±5.20)%, t=3.51,2.98,4.24, P < 0.01); But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 (t=2.01,2.36, P < 0.05), and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1(t=0.06,P > 0.05). CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.
文摘Objective: In this study, we aimed to investigate umbilical cord blood CD33 and erythropoietin (EPO) levels of pregnants with abnormal umbilical and uterine artery doppler waveforms and to compare with normal pregnancies. Materials and Methods: Total 40 pregnant women were included in this study. Of these 40 women, while 20 patients had abnormal umbilical and uterine artery doppler waveforms, the other 20 patients had normal umbilical and uterine artery doppler waveforms. After the delivery, blood samples were taken from umbilical artery of double clemped umbilical cord for blood gas parameters, EPO and CD33 levels. Sociodemographic findings, antepartum, intrapartum test results, labor and delivery characteristics and newborn examination results were recorded. Blood gas parameters, EPO and CD33 levels between groups were analyzed. Mann-Whitney U test and t-test were used as statistical methods. Results: There were no differences between parity, gestational ages and newborn weights of the groups. Cord blood CD33 and EPO levels of group with abnormal umbilical and uterine artery doppler waveforms were significantly higher than group with normal umbilical and uterine artery doppler waveforms (p < 0.01). Conclusion: Pathology on doppler screen shows to us a connection between chronic hypoxemia and abnormal on doppler screen. Preference of high blood CD33 levels for cord blood transplantation especially during last years can also be used with preference of cord blood with abnormal doppler findings.
基金This work was supported by the Finnish Funding Agency for Technology and Innovation(TEKES)the EVO Medical Research Fund of Finnish Red Cross Blood Service and The Finnish Glycoscience Graduate School.
文摘Umbilical cord blood(UCB)is an efficient and valuable source of hematopoietic stem cells(HSCs)for transplantation.In addition to HSCs it harbours low amounts of mesenchymal stem cells(MSCs).No single marker to identify cord blood-derived stem cells,or to indicate their multipotent phenotype,has been characterized so far.SSEA-3 and-4 are cell surface globoseries glycosphingolipid epitopes that are commonly used as markers for human embryonic stem cells,where SSEA-3 rapidly disappears when the cells start to differentiate.Lately SSEA-3 and-4 have also been observed in MSCs.As there is an ongoing discussion and variation of stem-cell markers between laboratories,we have now comprehensively characterized the expression of these epitopes in both the multipotent stem-cell types derived from UCB.We have performed complementary analysis using gene expression analysis,mass spectrometry and immunochemical methods,including both flow cytometry and immunofluoresence microscopy.SSEA-4,but not SSEA-3,was expressed on MSCs but absent from HSCs.Our findings indicate that SSEA-3 and/or-4 may not be optimal markers for multipotency in the case of stem cells derived from cord blood,as their expression may be altered by cell-culture conditions.
基金supplying HUCB and irradiating mice.This project was supported by project grant From Zhejiang Science Foundation (No.2006C23027).
文摘The effects of hematopoietic stem/progenitor cells(HSPCs)expanded in the two step coculture with human bone marrow mesenchymal stem cells(hMSCs)on the hematopoietic reconstruction of irradiated NOD/SCID mice were studied.Mononuclear cells(MNCs)were isolated from human umbilical cord blood(UCB)and cultured in the non-coculture scheme of rhSCF+rhG−CSF+rhMDGF combination and the coculture scheme of rhSCF+rhG−CSF+rhMDGF+hMSCs.Sublethally-irradiated NOD/SCID mice were transplanted with ex vivo expanded HSPCs with the dose of 8.5×10^(6) cells per mouse.After transplantation,the dynamics of WBC in the transplanted mice was measured periodically,and the Alu sequence fragment special for human in the transplanted mice was inspected by PCR.Results showed that the coculture scheme increased proliferation of UCB-derived HSPCs.After transplantation with expanded HSPCs,the population of WBC in the transplanted mice increased in 12 d and reached the first peak in 25 d,then showed the second increasing of WBC in 45~55 d.Expanded cells from the coculture scheme appeared to be favorable for the second increasing of WBC in the transplanted mice.After 85 d,the Alu sequence fragment was detected in the probability of 87.5%(7/8)for the non-coculture scheme and 88.9%(8/9)for the coculture scheme.
基金the National Key Research&Development Program of China(2016YFC1000400,2018YFC1002903).
文摘Objective:To evaluate the predictive ability of neonate condition through the traditional parameters and artery umbilical cord blood gas(aUCBG).Methods:A prospective cohort study was conducted in obstetrics and gynecology department between October 2017 and August 2018 at Tongji Hospital in Wuhan,China,and 360 aUCBG samples were collected.The average age of pregnant women was(29.50±4.42)years,range from 19 to 48 years old.The gestational age range from 28+4 weeks to 41+3 weeks at admission.Logistic regression and area under the curve(AUC)from Receiver operating characteristic curves were used to identify risk factors,such as,premature rupture of membranes(PROM),high blood pressure,premature delivery(PD),low 1-minute Apgar scores(Apgar 1),low 5-minute Apgar scores(Apgar 5),pH,base excess,bicarbonate,neonatal blood sugar(NBS),and so on,to predict neonatal condition and evaluate the predictive ability of traditional and aUCBG parameters.Results:In all cases,PROM,PD,Apgar 1,Apgar 5,pH,base excess,bicarbonate,total carbon dioxide,and neonatal blood sugar were risk factors and were associated with poor condition of neonate.Apgar 1 were an independent risk factor.Combined traditional and aUCBG parameters had higher AUC of 0.895(95%confidence interval(C/):0.830-0.960,P<0.001).In cesarean section subgroup,high blood pressure,PD,and Apgar 1 were risk factors and were associated with poor condition of neonate.Apgar 1 and low pH were the independent risk factors.Combined traditional and aUCBG parameters had highest AUC of 0.940(95%C/:0.886-0.993,P<0.001).In vaginal delivery subgroup,maternal age above 35 years,PROM,PD,Apgar 1,Apgar 5,and male newborn were risk factors and were associated with poor condition of neonate.Maternal age above 35 years was an independent risk factor.Combined traditional and aUCBG parameters had highest AUC of 0.897(95%Cl:0.828-0.965,P<0.001).For pregnant women without comorbidities and complications of pregnancy,aUCBG may not be necessat7.Conclusion:In high-risk pregnancies,especially lower Apgar scores,PD,and maternal age above 35-year old,aUCBG is recommended.Traditional parameters combined with aUCBG might increase the predicting ability of neonate condition.
基金supported by the National Natural Science Foundation of China,No.81273775
文摘OBJECTIVE: The aim of this study was to evaluate the effectiveness and safety of stem cell transplantation for spinal cord injury(SCI).DATA SOURCES: PubM ed, EMBASE, Cochrane, China National Knowledge Infrastructure, China Science and Technology Journal, Wanfang, and Sino Med databases were systematically searched by computer to select clinical randomized controlled trials using stem cell transplantation to treat SCI, published between each database initiation and July 2016. DATA SELECTION: Randomized controlled trials comparing stem cell transplantation with rehabilitation treatment for patients with SCI. Inclusion criteria:(1) Patients with SCI diagnosed according to the American Spinal Injury Association(ASIA) International standards for neurological classification of SCI;(2) patients with SCI who received only stem cell transplantation therapy or stem cell transplantation combined with rehabilitation therapy;(3) one or more of the following outcomes reported: outcomes concerning neurological function including sensory function and locomotor function, activities of daily living, urination functions, and severity of SCI or adverse effects. Studies comprising patients with complications, without full-text, and preclinical animal models were excluded. Quality of the included studies was evaluated using the Cochrane risk of bias assessment tool and Rev Man V5.3 software, provided by the Cochrane Collaboration, was used to perform statistical analysis. OUTCOME MEASURES: ASIA motor score, ASIA light touch score, ASIA pinprick score, ASIA impairment scale grading improvement rate, activities of daily living score, residual urine volume, and adverse events.RESULTS: Ten studies comprising 377 patients were included in the analysis and the overall risk of bias was relatively low level. Four studies did not detail how random sequences were generated, two studies did not clearly state the blinding outcome assessment, two studies lacked blinding outcome assessment, one study lacked follow-up information, and four studies carried out selective reporting. Compared with rehabilitation therapy, stem cell transplantation significantly increased the lower limb light touch score(odds ratio(OR) = 3.43, 95% confidence interval(CI): 0.01 – 6.86, P = 0.05), lower limb pinprick score(OR = 3.93, 95%CI: 0.74 – 7.12, P = 0.02), ASI grading rate(relative risk(RR) = 2.95, 95%CI: 1.64 – 5.29, P = 0.0003), and notably reduced residual urine volume(OR = –8.10, 95%CI: –15.09 to –1.10, P = 0.02). However, stem cell transplantation did not significantly improve motor score(OR = 1.89, 95%CI: –0.25 to 4.03, P = 0.08) or activities of daily living score(OR = 1.12, 95%CI: –1.17 to 4.04, P = 0.45). Furthermore, stem cell transplantation caused a high rate of mild adverse effects(RR = 14.49, 95%CI: 5.34 – 34.08, P < 0.00001); however, these were alleviated in a short time. CONCLUSION: Stem cell transplantation was determined to be an efficient and safe treatment for SCI and simultaneously improved sensory and bladder functions. Although associated minor and temporary adverse effects were observed with transplanted stem cells, spinal cord repair and axon remyelination were apparent. More randomized controlled trials with larger sample sizes and longer follow-up times are needed to further validate the effectiveness of stem cell transplantation in the treatment of SCI.
基金supported by the grant of Russian Science Foundation,No.16-15-00010(to RRI)supported by the Russian Government Program of Competitive Growth of Kazan Federal University。
文摘Despite emerging contemporary biotechnological methods such as gene-and stem cell-based therapy,there are no clinically established therapeutic strategies for neural regeneration after spinal cord injury.Our previous studies have demonstrated that transplantation of genetically engineered human umbilical cord blood mononuclear cells producing three recombinant therapeutic molecules,including vascular endothelial growth factor(VEGF),glial cell-line derived neurotrophic factor(GDNF),and neural cell adhesion molecule(NCAM)can improve morpho-functional recovery of injured spinal cord in rats and mini-pigs.To investigate the efficacy of human umbilical cord blood mononuclear cells-mediated triple-gene therapy combined with epidural electrical stimulation in the treatment of spinal cord injury,in this study,rats with moderate spinal cord contusion injury were intrathecally infused with human umbilical cord blood mononuclear cells expressing recombinant genes VEGF165,GDNF,NCAM1 at 4 hours after spinal cord injury.Three days after injury,epidural stimulations were given simultaneously above the lesion site at C5(to stimulate the cervical network related to forelimb functions)and below the lesion site at L2(to activate the central pattern generators)every other day for 4 weeks.Rats subjected to the combined treatment showed a limited functional improvement of the knee joint,high preservation of muscle fiber area in tibialis anterior muscle and increased H/M ratio in gastrocnemius muscle 30 days after spinal cord injury.However,beneficial cellular outcomes such as reduced apoptosis and increased sparing of the gray and white matters,and enhanced expression of heat shock and synaptic proteins were found in rats with spinal cord injury subjected to the combined epidural electrical stimulation with gene therapy.This study presents the first proof of principle study of combination of the multisite epidural electrical stimulation with ex vivo triple gene therapy(VEGF,GDNF and NCAM)for treatment of spinal cord injury in rat models.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.2.20.02.18)on February 20,2018.
文摘BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferating potential in kidney injury in mice.METHODS Human umbilical cord blood(UCB)-derived CD34+cells were incubated for one week in vasculogenic conditioning medium.Vasculogenic culture significantly increased the number of CD34+cells and their ability to form endothelial progenitor cell colony-forming units.Adenineinduced tubulointerstitial injury of the kidney was induced in immunodeficient non-obese diabetic/severe combined immunodeficiency mice,and cultured human UCB-CD34+cells were administered at a dose of 1×106/mouse on days 7,14,and 21 after the start of adenine diet.RESULTS Repetitive administration of cultured UCB-CD34+cells significantly improved the time-course of kidney dysfunction in the cell therapy group compared with that in the control group.Both interstitial fibrosis and tubular damage were significantly reduced in the cell therapy group compared with those in the control group(P<0.01).Microvasculature integrity was significantly preserved(P<0.01)and macrophage infiltration into kidney tissue was dramatically decreased in the cell therapy group compared with those in the control group(P<0.001).CONCLUSION Early intervention using human cultured CD34+cells significantly improved the progression of tubulointerstitial kidney injury.Repetitive administration of cultured human UCB-CD34+cells significantly improved tubulointerstitial damage in adenine-induced kidney injury in mice via vasculoprotective and anti-inflammatory effects.
基金funded by the subsidy allocated to Kazan Federal University for the state assignment#0671-2020-0058 in the sphere of scientific activities。
文摘Tay-Sachs disease and Sandhoff disease are severe hereditary neurodegenerative disorders caused by a deficiency ofβ-hexosaminidase A(HexA)enzyme,which results in the accumulation of GM2 gangliosides in the nervous system cells.In this work,we analyzed the efficacy and safety of cell-mediated gene therapy for Sandhoff disease and Sandhoff disease using a bicistronic lentiviral vector encoding cDNA of HexAα-andβ-subunit genes separated by the nucleotide sequence of a P2A peptide(HEXA-HEXB).The functionality of the bicistronic construct containing the HEXA-HEXB genetic cassette was analyzed in a culture of HEK293T cells and human umbilical cord blood mononuclear cells(hUCBMCs).Our results showed that the enzymatic activity of HexA in the conditioned medium harvested from genetically modified HEK293T-HEXA-HEXB and hUCBMCs-HEXA-HEXB was increased by 23 and 8 times,respectively,compared with the conditioned medium of native cells.Western blot analysis showed that hUCBMCs-HEXA-HEXB secreted both completely separated HEXA and HEXB proteins,and an uncleaved protein containing HEXA+HEXB linked by the P2A peptide.Intravenous injection of genetically modified hUCBMCs-HEXA-HEXB to laboratory Wistar rats was carried out,and the HexA enzymatic activity in the blood plasma of experimental animals,as well as the number of live cells of immune system organs(spleen,thymus,bone marrow,lymph nodes)were determined.A significant increase in the enzymatic activity of HexA in the blood plasma of laboratory rats on days 6 and 9(by 2.5 and 3 times,respectively)after the administration of hUCBMCsHEXA-HEXB was shown.At the same time,the number of live cells in the studied organs remained unchanged.Thus,the functionality of the bicistronic genetic construct encoding cDNA of the HEXA and HEXB genes separated by the nucleotide sequence of the P2A peptide was shown in vitro and in vivo.We hypothesize that due to the natural ability of hUCBMCs to overcome biological barriers,such a strategy can restore the activity of the missing enzyme in the central nervous system of patients with GM2 gangliosidoses.Based on the obtained data,it can be concluded that intravenous administration of hUCBMCs with HexA overexpression is a promising method of the therapy for GM2 gangliosidoses.The animal protocol was approved by the Animal Ethics Committee of the Kazan Federal University(No.23)on June 30,2020.
文摘Autism spectrum disorder(ASD)is a complex neurodevelopmental condition characterized by impairments in social communication,abnormal to absent verbal communication,the presence of repetitive stereotypic verbal and nonverbal behaviors and restricted interests,with onset early in life.We showed cognitive and behavioral characteristics of ASD by impairment of communication,cognition,perception,motor skills,executive function,theory of mind and emotion control.Recently,pathogenesis of immune pathology in the brains of individuals with ASD has been focused.New therapeutic approaches in the viewpoints of immune modulation and microglial function are logical for novel treatments for individuals with ASD.Cell therapies such as umbilical cord blood cells and mesenchymal stromal cells for ASD will be a challenging and encouraging field in the future clinical study with progress of biotechnological science.
文摘Diabetic patients often exhibit delayed or incomplete progress in the healing of acute wounds,owing to poor blood perfusion.Platelet-rich plasma(PRP)has attracted much attention as a means to improve wound healing,because it contains high growth factor concentrations.However,the burst-like release of PRP growth factors results in a short halflife of these therapeutic proteins,thus greatly limiting the therapeutic effect.In this study,we prepared PRP from human umbilical cord blood and developed an in situ photocrosslinkable PRP hydrogel glue(HNPRP)by adding a photoresponsive hyaluronic acid(HA-NB)into PRP.The HNPRP hydrogel allowed for controlled release of plateletderived growth factor-BB(PDGF-BB)and transforming growth factor-β(TGF-β)for up to 28 days.In vitro cell culture showed that HNPRP promoted migration of fibroblasts and keratinocytes as well as PRP and did not reveal the advantages of HNPRP.However,in a diabetic rat skin wound model,HNPRP treatment promoted faster wound closure.Furthermore,the HNPRP group,compared with the control,PRP and hydrogel only groups,showed significantly greater re-epithelialization and numbers of both newly formed and mature blood vessels.The HNPRP group also displayed higher collagen formation than did the control group.In conclusion,HNPRP enhances angiogenesis and skin regeneration and consequently achieves faster wound healing,thus extending its potential for clinical applications to treat diabetic skin wounds.
