Objective:To study methods for inducing CD34+ cells of human bone marrow to differentiate into T cells in vitro to provide theory and method basics for the investigation of activity of T cells derived from psoriatic bone marrow CD34+ cells and establish a technological platform to investigate T lymphopoiesis activity of hematopoietic cells.Methods:Bone marrow CD34+ hematopoietic progenitor cells were isolated by immunomagnetic cell selection and induced differentiate into T cells in the bone in the marrow and thymic stromal microenvironment.Immunfluorescence dying method and flow cytometry analysis were performed to detect CD1-CD3+ cells,CD3+CD4+CD8-cells and CD3+CD4-CD8+ cells dynamically.Results:In the first week,the non-adhension cells were composed mostly of immature CD1+CD3-cells and CD1+CD3+ cells and small proportion of mature CD1-CD3+ cells.In the following analysis,the proportion of immature cells rapidly decreased and CD1-CD3+ cells increased.After 1 week culture,CD4+CD8+ double positive T cells and a small population CD4+CD8-and CD4-CD8+ could be detected among the CD3+ cells.In the following culture,the proportion of CD4+CD8+ double positive T cells decreased significantly and single positive T cells increased gradually.However,small proportion of mature T cells could be detected in the early stage and cann't be found after 4 weeks in the culture system without thymic stromal cells.Conclusion:Mature single positive T cells can develop from CD34+ hematopoietic progenitor cells in the bone marrow and thymic stromal microenvironment and the thymic stromal cells are vital for T lymphopoiesis.

Objective To investigate the mechanism of abnormal epidermal proliferation induced by psoriatic T lymphocytes.Methods Skin organ culture was established with psoriatic T cells mixed with epi-dermal cells.The expressions of c-myc,bcl-2and p53protein were examined with immunohistochemical method in epidermis before culture,on the3rd day and the6th day after culture.Results Significant up-regulation of c-myc and p53protein was found in psoriatic lesions,but bcl-2protein expression was rarely observed.The expressions of those proteins were normal in non-lesional psoriatic skin.The p53protein ex-pression was increased in normal skin and non-lesional psoriatic skin on the3rd day after culture with psori-atic T cells,and c-myc protein expression was enhanced while bcl-2was decreased on the6th day of co-culture.There was no significant difference of those proteins' expression between normal skin and non-lesion-al psoriatic skin stimulated by psoriatic T cells.Conclusions The abnormal expressions of c-myc,bcl-2and p53protein play an important role in abnormal epidermal proliferation and differentiation in psoriasis.Psoriatic T lymphocytes can influence c-myc,bcl-2and p53protein expression in normal skin and non-le-sional psoriatic skin.Pathogenic T cells rather than keratinocytes might be vital for initiation of psoriasis.

Objective:To compare the seeding efficiency and graft versus host disease (GVHD) of severe combined immunodeficient (SCID) mice transplanted with human hematopoietic cells intraperitoneally (ip) and intravenously (iv) and to explore the method to set up psoriatic animal model by xenogeneic bone marrow transplantation.Methods:Normal human bone marrow mononuclear cells (BMMNC) were separated by density gradient centrifugation and BMMNC (4?10~7) were injected into lethally irradiated SCID mice by ip or iv injection. GVHD symptom and periphery blood white blood cell resuming dynamics in mice were observed after xenotransplantation and flow cytometry was performed to detect human source CD45~+ cells proportion in periphery blood and bone marrow of mice.Results:The mice transplanted by tail intravenous injection presented obvious GVHD symptoms promptly 2 weeks after transplanting and only one mouse survived in 12 weeks. Among the mice received tail intravenous injection and dealed with cydosporin(CsA) and methotrexate(MTX),some of the mice showed slight GVHD symptom and survival rate was 80%(8/10) in 12 weeks. Slight GVHD symptoms appeared after human bone marrow transplantation by intraperitoneal injection and then most of mice returned to the normal and the survival rate was 70%(7/10) in 12 weeks. The periphery blood white blood cells resuming dynamics, CD45~+ cell proportion of periphery blood and bone marrow after transplantation show no significant difference between the groups transplanted by intravenous and intraperitoneal injection.Conclusion:Human hematopoietic cells could home to bone marrow in SCID mice and result in hematopoietic reconstitution. The transplantation method by intraperitoneal injection, which showed efficient seeding capability, can be used to both bone-marrow transplantation and reducing GVHD.

BACKGROUND: It remains disputed whether bone filling bag vertebroplasty and percutaneous kyphoplasty have different treatment efficacy in the treatment of thoracolumbar osteoporotic compression fractures. OBJECTIVE: To systematically analyze the efficacy and safety of bone filling bag vertebroplasty and percutaneous kyphoplasty in the treatment of thoracolumbar osteoporotic compression fractures. METHODS: A computer-based online search of CNKI, Wanfang, VIP, CBM, EMBASE, MEDLINE, and Cochrane libraries was performed to retrieve randomized controlled trial studies regarding bone filling bag vertebroplasty and percutaneous kyphoplasty in the treatment of thoracolumbar osteoporotic compression fractures published before February 2019. Two researchers independently conducted literature screening and data extraction. According to the Cochrane Collaboration Network standard, the quality of the randomized controlled trial studies was evaluated one by one. The studies that met the inclusion criteria were analyzed using the RevMan5. 3 software. RESULTS AND CONCLUSION: Six randomized controlled trial studies were included. A total of 517 patients were included in the final analysis. Among them, 257 patients received bone filling bag vertebroplasty and 260 patients received percutaneous kyphoplasty. Meta-analysis showed that there were no significant differences in postoperative Visual Analogy Score (MD=0. 00, 95%CI: -0. 09-0. 10, P=0. 94), vertebral height recovery (SMD=0. 11, 95%CI: -0. 26-0. 48, P=0. 57), and Oswestry Disability Index (MD=1. 47, 95%CI: -0. 45-3. 39, P=0. 13) between these two surgical procedures. But postoperative Cobb angle (MD=-1. 08, 95%CI: -1. 47 to -0. 70, P < 0. 000 01) and bone cement leakage rate (RR=0. 24, 95%CI: 0. 13-0. 45, P < 0. 000 01) were significantly different between these two surgical procedures. Bone filling bag vertebroplasty exhibits significant advantages in improving postoperative Cobb angle and reducing bone cement over percutaneous kyphoplasty. These two surgical procedures have similar clinical outcomes such as postoperative Visual Analogy Score, vertebral height recovery, and Oswestry Disability Index. Therefore, a large number of high-quality multicenter randomized controlled trials are needed to provide more evidence.

Objective:To reveal the relation between RUNX1 and hematopoietic cells by examination of the expression of RUNX1 and its target gene SLC9A3R1 in bone marrow CD34+ cells from patients with psoriasis,as well as the RUNX1 linkage locus between SLC9A3R1 and NAT9.Methods:Bone marrow CD34+ cells were isolated from psoriatic patients and normal persons by immunomagnetic cell selection.Expression of mRNA for RUNX1 and SLC9A3R1 were analyzed using reverse transcriptase-polymerase chain reaction(RT-PCR),the RUNX1 linkage locus between SLC9A3R1 and NAT9 was detected.Results:The positive frequency of RUNX1 in bone marrow CD34+ cells from patients with psoriasis was lower than in normal controls(P

Objective To investigate the effects of psoriatic keratinocytes on the expression of CD25 and CD69 in T lymphocytes. Methods Keratinocytes were isolated from the biopsy samples resected from the lesions and adjacent non-lesional area of 10 patients with psoriasis, and cultured in 5% CO2 at 37 ℃ in 24-well plates. Density gradient centrifugalization and glass adherence method were applied to detach peripheral blood mononuclear cells (PBMC) and peripheral blood T lymphocytes (PBTL) from anticoagulant blood samples of the same 10 psoriatic patients and 10 normal controls. PBMCs of 1×105/well were added to the wells containing cultured keratinocytcs of 1×105/well, then gamma rays were used to inactivate these cells. Following that, PBTLs of 1×106/well were inoculated into the 24-well plate containing inactivated keratinocytes and PBMCs, and cultured in 5% CO2 at 37 ℃. Those PBTLs cultured without the presence of keratinocytes or PBMCs served as the natural growth control. Three days later, flow cytometry was performed to detect the expression of CD25 and CD69 in PBTLs. Results There was a significant increase in the expression of CD25 and CD69 in psoriatic PBTLs cocultured with lesional kcratinocytes compared with those cocultured with non-lesional keratinocytes and natural psoriatic controls. Also, the expression of CD25 and CD69 was increased in normal PBTLs cocultured with lesional or non-lesional keratinocytes of psoriatc patients than those in the natural normal controls. No significant differenco was observed in the expression of CD25 or CD69 between psoriatic PBTLs cocultured with non-lesional keratinocytes and natural psoriatic PBTLs, or between the normal PBTLs cocultrred with lesional keratinocytes and those with non-lesienal keratinocytes (P>0.05). Conclusions Psoriatic keratinocytes may act as an autoantigen to trigger autoimmune response and eventually lead to a chronic local inflammation in patients with psoriasis.

Objective To investigate effect of respiratory training on respiratory function recovery following thoracotomy. Method Respiratory muscle training, productive drainage, exercise and respiratory function training, and appropriate rehabilitation were performed on 216 patients underwent thoracotomy. Result 206 patients showed favorable recovery of respiratory function,7 showed respiratory dysfunction, and 1 developed serious complication. Conclusion Respiratory training after thoracotomy significantly improve respiratory function.

Objective To study the influence of culture supernatant of psoriatic peripheral blood mononuclear cells (PBMCs) on the colony-forming of bone marrow-derived hematopoietic stem cells and progenitor cells. Methods Bone marrow-derived mononuclear cells were separated by density gradient centrifugation. Methylcellulose semi-solid culture medium was used to culture the bone marrow mononuclear cells in culture systems. The PBMC culture supernatant from psoriatic patients or normal controls were added to the culture, to observe their influence on the marrow-derived high proliferative potential colony forming cells (HPP-CFCs), erythroid and granulocyte-macrophage colony forming units (CFUs-E and CFUs-GM) of bone marrow hematopoietic stem/progenitor cells from healthy individuals. Results The values of HPP-CFCs, CFUs-E and CFUs-GM were significantly lower in the bone marrow cells stimulated by the supernatant of cultured psoriatic PBMCs than those stimulated by the supernatant of cultured normal PBMCs and those in the spontaneous proliferation group (P 0.05). Conclusions Psoriatic PBMCs have specific biological activity and can inhibit the colony forming of bone marrow hematopoietic cells from healthy individuals.

Objective To study the in vitro activity of T cells differentiated from bone marrow CD34+ cells of psoriatic patients with family histories. Methods Bone marrow CD34+ cells were isolated from psoriatic patients with family history and normal persons by immunomagnetic cell selection and induced to differentiate to T cells in thymic stromal microenvironment in vitro. CD3+ T cells were obtained by CD3+ cell selection system and the proportions of CD4+CD8- and CD4-CD8+ cells were measured by flow cytometry. The proliferation activity of T cells was measured by MTT colorimetry in both spontaneous proliferating group and superantigen (SAg)-stimulated group. And the levels of IL-4, IL-8 and IFN-? were quantified by enzyme-linked immunosorbent assay in culture supernatant. Results Mature CD4+CD8- cells and CD4-CD8+ cells were detected from CD3+ cells, which were derived from bone marrow CD34+ cells. When the psoriatic patients and normal controls were compared, there was no significant difference in the proportion of CD4+CD8- cells or that of CD4-CD8+ cells in CD3+ cells. The proliferation activity of T cells was significantly higher in the psoriatic patients than that in the normal controls. There was no statistical difference of IL-4?IL-8 or IFN-? levels in the supernatant of T cells between the spontaneous group and the normal control. However, the IFN-? and IL-8 levels in the supernatant of T cells were higher in the psoriatic patients than those in the normal controls after the SAg stimulation. Conclusion The abnormal activity of peripheral blood T cells from psoriatic patients with family history may be related to the bone marrow hematopoietic cells.

Objective To investigate the effect of preoperative acute hypervolemic hemodilution with 6% hydroxyethyl starch 130/0.4(HES130/0.4)and polygeline on hemodynamic response and variance regularity of hemodynamic response in open-and close-thoracic cavity in undergoing thoracotomy patients. Methods Sixty patients undergoing thoracotomy,ASA physical status ⅠorⅡ,aged 20~65 yeansold,weighing 45~70kg,were randomly divided three group:HES130/0.4 group ( group A,n=20) ,Polygeline group ( group B,n=20) and Ringer's solution group (group C,n=20).They were respectively infused 20ml/ (kg?h) with HES130/0.4,polygeline and Ringer's solution in one hour before infuseing followed Ringer's 10ml/ (kg?h). The Blood Pressure (BP) and heart rate (HR) were recorded.RPP was calculated before induction(baseline),5min before and 10 min after open thoracic cavity,5min before and 10min after close thoracic cavity. And central venous pressure (CVP) was measured at immediately after deep venous puncture ,5min before and 10min after open thoracic cavity,5min before and 10min after close thoracic cavity. Results The changes of hemodynamics in Ringer's solution group in operation were obvious as compared with baseline before operation.HES130/0.4 and polygeline were relatively stable.Two groups had no significant difference.The CVP of HES130/0.4 and polygeline groups increased obviously before and after open thoracic cavity,but the blood pressure was relatively stable.Only systolic blood pressure increased obviously as compared with Ringer's solution group at 5min before open thoracic cavity. At 10min after open thoracic cavity,HR and CVP in three groups increased obviously as compared with 5 min before open thoracic cavity.At 5 min before close thoracic cavity,blood pressure and CVP in three groups decreased obviously as compared with 10 min after open thoracic cavity. Conclusion The preoperative acute hypervolemic hemodilution with HES 130/ 0.4 and polygeline in undergoing thoracotomy patients can keep hemodynamics more stable and improve tolerance of patients to withstand hypovolemics.