BACKGROUND:Separating umbilical cord blood stem cells using tubes has low efficiency,and microbial contamination easily occurs during this process,therefore,safety cannot be ensured in clinical application.lt is urgent to find a method for separating umbilical cord blood stem cells to treat femoral head necrosis.OBJECTIVE:To establish a high efficient,safe,and clinically valuable method to separate umbilical cord blood stem cells.DESIGN,TIME AND SETTING:A self-control experiment was performed at the First Department of Surgery,Zhengzhou Second People's Hospital,Institute of Blood Constituent Application,Henan Red Cross Blood Centre between February 2006 and August 2007.PARTICIPANTS:Eight male patients with femoral head necrosis,averaging 40.6 years of age,were included in this study.Of these patients,4 had the history of hormone application.An average of 90 mL umbilical cord blood was harvested from each healthy normal full term neonate from Maternal and Children Health Care Hospital of Zhengzhou City.The quadruple bags used for separating umbilical cord blood stem cells consisted of 1 main bag,1 empty bag,and 2 physiological saline bags,provided by Shandong Weigao Holding,China.METHODS:Within 6 hours after collection,umbilical cord blood was centrifuged in the empty bag of quadruple bag,which was connected with an aseptic filling machine.After centrifugation,partial blood plasma was discarded,and the remaining erythrocytes were thoroughly mixed by adding hetastarch.Five minutes later,the mixture was diluted with physical saline at 1:1.Umbilical cord blood was slowly added into the main bag (at 1:1),in which,human lymphocyte separating medium was pre-added.After cantrifugation,the upper layer of solution,i.e.,monocyte-rich solution,was transferred into another empty bag.Within 24hours of preservation,after suspension with umbilical blood plasma,umbilical cord monocytes were transfused into patients with femoral head necrosis via superficial vein on the hand back,monocytes≥1×108/portion,2 portions once.There were three treatment courses,each involving three transfusion sessions,one session every 4 days,and a 2-3-month interval between two treatment courses.MAIN OUTCOME MEASURES:Cell recovery rate and cell viability of umbilical cord blood monocytes and improvements in clinical symptoms.RESULTS:The separation of quadruple bags could obtain umbilical cord blood monocytes with high recovery rate.Furthermore,microbial contamination hardly occurred in the process of separation.Hip joint pain relieved or disappeared to different extents in all 8 patients,with an effective rate of 100%.Abduction and internal rotation of hip joint,ambulation distance,and gait were markedly improved.At 6 months after cell transplantation,5 patients presented with changed bone density in femoral head necrosis regions,2 showed normal femoral head morphology,and the remaining 1 exhibited no obvious changes.Joint effusion was reduced or disappeared in 12 hips.Magnetic resonance images showed that femoral head morphology had been improved in various degrees in 9 hips,but no changes in 3 hips.No complications,fever,or allergies occurred during and after cell transplantation.CONCLUSION:The method of separating stem cells from umbilical cord blood in junction with aseptic interface technology is highly effective,safe,and clinically valuable.Multiple intravenous transfusions of umbilical cord blood stem cells provide a novel approach for systemic treatment of femoral head necrosis.

BACKGROUND:Umbilical cord blood as a source of hematopoietic stem cel s has become a hot topic. Adjuvant therapy of umbilical cord blood transfusion has been used in China to correct renal anemia of uremia patients. OBJECTIVE:To evaluate the efficacy and safety of umbilical cord blood mesenchymal stem cel s in the treatment of renal anemia, and to compare with the effects of different sources of mesenchymal stem cel s with different transplantation methods on the treatment of kidney diseases. METHODS:A retrospective analysis was conducted on six renal anemia patients in the Department of Outpatient, Henan Red Cross Blood Center between January 2010 and December 2012. The experiment was approved by Medical Ethics Committee, six patients were informed consent for treatment programs, and parturients and their families have signed the informed consent. The newborns umbilical cord blood of 80-140 mL were col ected with closed sterile plastic blood bags, and then the separated umbilical cord blood mesenchymal stem cel s were transfused into the renal anemia patients through the superficial vein in the back of the hand with the number of≥1×108/copy, two copies per time, and re-transfused after 4 days, a total of three times. The blood hematocrit, hemoglobin, red blood cel s in urine, renal blood flow changes were observed before and after treatment. RESULTS AND CONCLUSION:The hemoglobin, blood hematocrit, red blood cel s in urine and renal blood flow were significantly increased before and after treatment (P<0.05). Transfusion of multiple copies of umbilical cord blood mesenchymal stem cel s through the superficial vein in the back of the hand is convenient and safe, which is considered as a new method for the treatment of renal anemia. But there are certain limitation in the data and the conclusion as the clinical research was designed as self-control, so further confirm is needed.

Objective To investigate whether the implanted spike wave electrical stimulation (ES) can improve survival of mesenchymal stem cells (MSCs) after transplantation in spinal cord injury (SCI) rats. MethodsIn the study,60 male SD rats were used to prepare the SCI induced by a digital weight drop apparatus.The MSCs from each rat' s femurs were collected and then labeled by Bromodeoxyuridine (Brdu) seven days after SCI.The rats were randomly assigned into four groups,ie,group A (receiving a transplantation of MSCs only),group B (receiving electro-stimulant therapy only),group C ( receiving a transplation of MSCs and electro-stimulant therapy) and the control group ( receiving a single injection of phosphate buffer solution,PBS).Functional status was assessed regularly before and after SCI by using the Basso-Beattie-Bresnahan (BBB) locomotor rating score and somatosensory evoked potential (SEP).The recovery of spinal cord was checked by diffusion tensor imaging (DTI) and the survivorship of the labeled MSCs was observed with immunohistochemical method. Results BBB score of group C demonstrated significant difference in comparison with that of the other three groups ( P ＜ 0.05 )and was significantly higher than that of the control group (P ＜0.01 ) four weeks after surgery,which indicated an obvious functional improvement of the extremity mobility in group C.At the 8th week postoperatively,SEP study displayed significant difference in group C from the other three groups in terms of latency and wave amplitude (P ＜0.05 ),indicating the fast neurofunctional recovery of the spinal cord in group C.MR images of the spinal cords by DTI showed the same outcome.Immunohistochemistry confirmed a large number of the labeled MSCs positive cells in the lesion site of group C.ConclusionTheimplanted spike wave ES promotes the bioactivity and survival of MSCs and improves the therapeutic effect in combination with MSCs transplantation.

BACKGROUND:Bone marrow mesenchymal stem cells have low immunogenicity and immunomodulatory effect,but there are seldom reports concerning the immunomodulatory effect of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord and its mechanims.OBJECTIVE:To investigate the immunomodulatory effects and mechanisms of Wharton's jelly-derived mesenchymal stem cells of human umbilical cord on varient peripheral blood T lymphocytes.METHODS:Mesenchymal stem cells were isolateded from Wharton's jelly of human umbilical cord by tissue culture.T lymphocytes from human peripheral blood were stimulated by phytohemagglutinin and co-cultured with umbilical cord Wharton's jelly-derived mesenchymal stem cells and umbilical cord Wharton's jelly-derived mesenchymal stem cells supernatant respectively to measure A value following 72 hours of coculture using multifunctional microplate reader.Expression of cytokines including transforming growth factor-beta 1(TGF-β1)and interferon-y(IFN-γ)was evaluated by enzyme-labeled immunosorbent assay.RESULTS AND CONCLUSION:Wharton's jelly-derived mesenchymal stem cells could inhibite the proliferation of T lymphocytes induced by phytohemagglutinin.The proliferation inhibition rate was 56%(P＜0.01).Wharton's jelly-derived mesenchymal stem cells supernatant also had inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin,in a dose-dependent fashion.The proliferation inhibition rates were 8.3% and 27% respectively in the 50% Wharton's jelly-derived mesenchymal stem cells supernatant and 100% Wharton's jelly-derived mesenchymal stem cells supematant groups(P＜0.05).Wharton's jelly-derived mesenchymal stem cells significantly decreased γ-interferon secrted from T-lymphocytes(P＜0.05).The secretion of TGF-β1 was lower in the coculture of Wharton's jelly-derived mesenchymal stem cells and T lymphocytes group than Wharton's jelly-derived mesenchymal stem cells alone group(P＜0.05).These indicated that Wharton's jelly-derived mesenchymal stem cells and Wharton's jelly-derived mesenchymal stem cells supernatant have inhibitory effects on proliferation of T lymphocytes induced by phytohemagglutinin.The mechanims may be associated with cell contant and inhibition of v-interferon secrted from T-lymphocytes.

Objective:To investigate the mechanism of programmed death 1(PD-1)/ programmed death ligand 1(PD-L1) signaling pathway and its feasibility as a potential therapeutic target and prognostic predictor by detecting the expressions, of PD-1 and PD-L1 in bone marrow mononuclear cells of children with acute lymphoblastic leukemia (ALL), and to provide new ideas for the diagnosis and treatment of ALL as well.Methods:Bone marrow samples were collected from 59 children with ALL in the First Affiliated Hospital of Zhengzhou University from September 2018 to July 2019.Flow cytometry was applied to detect the expression of PD-1 and PD-L1 in bone marrow mononuclear cells in 59 ALL patients, including 47 newly-diagnosed ALL patients and 12 relapsed ALL patients, respectively, at initial diagnosis, after induction therapy and early intensive treatment.Their relevant clinical data were collected and compared with the bone marrow specimens of 12 children suffering from non-malignant blood diseases as the control group of the same hospital during the same period.Results:There was no significant difference in the expression of PD-1 in the bone marrow mononuclear cells of the primary diagnosis group, recurrence group and control group ( H=2.402, P>0.05). The expression of PD-L1 in the relapsed and refractory group [(7.32±3.60)%] and the newly diagnosed group [(3.18±2.37)%] was higher than that in the control group [(0.84±0.39)%], and the differences were statistically significant ( H= 28.048, P<0.05). In the initial treatment group, the expression of PD-L1 in the bone marrow mononuclear cells was the strongest expression before treatment ( B=1.293), followed by after induction treatment ( B=0.036) and after early intensive treatment ( B=0.000), suggesting that there was a downward trend as the continued treatment.The expression of PD-L1 was the weakest expression in the low-risk group ( B=-3.912) than in the medium-risk group ( B=-3.595) and high-risk group ( B=0.000), revealing that the expression of PD-L1 is related to the risk grades of ALL.The higher the risk rating is, the higher the PD-L1 protein expression is. Conclusions:The high expression of PD-L1 may be involved in the pathogenesis and be used as an adverse predictor of ALL childhood and an evaluation index of chemotherapy efficacy.PD-1 / PD-L1 signaling pathway may be a potential therapeutic target of ALL childhood.

【Objective】 To analyze the causes of immune hemolytic transfusion reaction in one case, identify related antibodies, and explore transfusion compatibility testing. 【Methods】 ABO/Rh blood group identification, unexpected antibody identification of serum and diffusion fluid, direct antiglobulin test(DAT) and cross matching were conducted by saline method and/or microcolumn gel method. 【Results】 The patient′s blood group was O, and Rh phenotype was identified as DCCee. The DAT was negative, with strong anti-E antibody and weak anti-c antibody detected. Acute hemolytic transfusion reaction occurred in the patient after the last transfusion. 【Conclusion】 Currently, immune hemolytic transfusion reaction in China are mainly caused by Rh blood group system antibodies. The absence of unexpected antibody screening before blood transfusion and the weak anti-c antibody which resulted in missed detection of non compatibility in cross matching led to acute hemolytic transfusion reaction. It is recommended to conduct unexpected antibody screening before blood transfusion, and to collect blood sample for testing as soon as possible to improve the accuracy of DAT when acute hemolytic transfusion reaction is suspected.

The etiology and pathogenesis of sleep obstructive apnea hypopnea syndrome (OSAHS) is not yet definitive, evidence shows that the dysfunction of pharyngeal nerve and the atonia of the muscle innervated by these nerve could play an important role in the progress of OSAHS. Dopamine is a neurotransmitter in the central nervous system which significantly affects the sleep-awake regulation. So far mounting evidence shows that dopamine has a potential role in the modulation of hypoglossal nucleus. The progress of dopamine in obstructive sleep apnea hypopnea syndrome is reviewed in this article.