Objective To investigate the effect of tumor associated glycoprotein-72 （TAG-72） redirected T lymphocytes on breast cancer cells. Methods Peripheral blood mononuclear cells （PBMCs）were isolated from healthy volunteers. The recombinant vector anti-TAG-72-scFv-CD3ζ-pcDNA 3.0 were transfected into PBMCs by lipofectamineTM2000 （transfection group）， PBMCs transfected with plasmid pcDNA 3.0 as control group. MCF-7 and Bcap37 cells were cocultivated with PBMCs of transfection group and control group， respectively， and antitumor response of G1 block was observed. Results G1 block rate of MCF-7 cells in transfection group was （82.3±6.9）%， which was significantly higher than that in control group 〔（43.4±3.9）%， P＜0.05〕. G1 block rate of Bcap37 cells in transfection group was （51.3±4.7）%， and not differed from that in control group 〔（45.6±2.5）%， P＞0.05〕. Conclusion TAG-72 redirected T lymphocytes can inhibit the cell proliferation of TAG-72 positive breast cancer cells， and it may provide valuable tools for the cellular immunotherapy.
Objective To study the expression of NLRP3 inflammasome and its downstream inflammatory factors in patients with chronic obstructive pulmonary disease (COPD) and healthy controls, and to reveal the effect and significance of NLRP3 inflammasome in the pathogenesis of COPD. Methods Forty patients with acute exacerbation COPD (AECOPD) who were hospitalized from November 2016 to May 2017 were recruited in the AECOPD group, and recruited in the stable COPD group when they entered the stable stage. Forty healthy individuals were recruited in the control group. General information and peripheral blood were collected from each subject. The levels of NLRP3 mRNA and caspase-1 mRNA in peripheral blood mononuclear cells were measured by real-time PCR. The levels of IL-18 and IL-1β were measured by enzyme-linked immunosorbent assay. Results The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the stable COPD group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/ml vs. 1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/ml, all P<0.05] . The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the control group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/mlvs. 1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. The levels of NLRP3 mRNA, IL-18 and IL-1β were significantly higher in the stable COPD group than the control group [1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/mlvs. (1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. Correlation analysis showed that the plasma IL-18 level was positive correlated with leukocyte count and neutrophil percentage in the AECOPD group (r=0.372, P<0.05;r=0.386, P<0.05). The expression of NLRP3 mRNA in the AECOPD group and stable COPD group were positively correlated with the CAT score (r=0.387, P<0.05;r=0.399, P<0.05) . Conclusion NLRP3 inflammasome is involved in the inflammatory response in COPD patients.
ObjectiveTo assess the long-term effectiveness and safety of autologous bone marrow mononuclear cells (BM-MNC) transplantation in the treatment of critical diabetic lower arteriosclerosis obliterans (ASO). MethodsBetween January 2007 and January 2010, 61 patients with critical diabetic lower ASO were treated with standard medical therapies in 29 cases (control group) or with standard medical therapies and autologous BM-MNC transplantation in 32 cases (treatment group). There was no significant difference in gender, age, disease duration, Fontatine stage, glucose (GLU), triglyceride (TG), total cholesterol (CHOL), low-density lipoprotein-cholesterol (LDL-C), hemoglobin A1c (HbA1c), systolic blood pressure (SBP), and diastolic blood pressure (DBP) between 2 groups (P>0.05). The endpoints were overall survival (OS) and amputation-free survival (AFS). The risk indexes for ASO were observed and compared between 2 groups before and after treatments. ResultsThe patients were followed up 2-36 months, and no malignant tumor occurred. The OS rate, OS time, AFS rate, and AFS time were 82.76% (24/29), (32.31±9.08) months, 37.50% (9/24), and (21.28±13.35) months in the control group and were 78.13% (25/32), (32.47±6.96) months, 68.00% (17/25), and (28.38±9.48) months in the treatment group;all indexes showed no significant differences (P>0.05). OS rate, OS time, AFS rate, and AFS time showed no significant differences between 2 groups at the other time (P>0.05) except AFS time at 1 year, which was significantly short in the control group than the treatment group (t=2.806, P=0.007). At the endpoint of follow-up, the indexes of GLU, TG, CHOL, LDL-C, HbA1c, SBP, and DBP showed no significant differences between before and after treatments and between 2 groups (P>0.05) in 49 survival patients (24 in control group and 25 in treatment group). ConclusionAutologous BM-MNC transplantation is safe and effective in the treatment of critical diabetic lower ASO, which can significantly improve AFS rate and prolong AFS time with no risks.
【摘要】 目的 检测B细胞成熟抗原（BCMA）mRNA在系统性红斑狼疮（SLE）患者外周血单个核细胞（PBMC）的表达水平，探讨BCMA在SLE发病中的意义。 方法 纳入2006年1-11月收治的36例SLE患者，同期17例健康志愿者作为对照组，采用半定量RT-PCR法检测外周血单个核细胞中BCMA mRNA的表达，并与SLE疾病活动指数（SLEDAI）进行相关性分析。 结果 SLE患者组BCMA mRNA表达水平（0.598±0.230）均明显高于正常对照组（0.411±0.309）（Plt;0.05）。SLE患者BCMA mRNA表达水平与SLEDAI评分无相关性（P=0.590）。 结论 SLE患者BCMA mRNA表达水平的增高，可能在SLE的发病机制中具有一定的作用。【Abstract】 Objective To detect the mRNA expression of B-cell maturation antigen (BCMA) in peripheral blood mononuclear cells (PBMC) in patients with systemic lupus erythematosus (SLE), and explore the role of BCMA in the pathogenesis of SLE. Methods From January 2006 to November 2006 the expression of BCMA mRNA in PBMC of 36 patients with SLE and 17 normal controls were measured by half-quantitative RT-PCR. The linear correlation between the expression of BCMA mRNA and SLE disease activity index (SLEDAI) was assessed. Results The level of BCMA mRNA (0.598±0.230) in PBMC significantly increased in SLE patients compared with that in the normal controls (0.411±0.309) (Plt;0.05). The expression of BCMA mRNA in SLE patients showed no correlation with SLEDAI score (P=0.590). Conclusion The results suggest that the expression of BCMA mRNA might play an important role in the pathogenesis of SLE.
ObjectiveTo systematically review the efficacy and safety of autologous mononuclear cells transplantation in osteonecrosis of the femoral head.MethodsPubMed, EMbase and The Cochrane Library were electronically searched to collect randomized and non-randomized controlled trials on autologous mononuclear cells transplantation for osteonecrosis of the femoral head from inception to July 31th, 2020. Two reviewers independently screened literatures, extracted data and assessed risk of bias of included studies. Meta-analysis was then performed using RevMan 5.4 software.ResultsA total of 17 studies involving 645 hips in mononuclear cells group and 557 hips in cell-free group were included. The results of meta-analysis showed that compared with cell-free therapy, mononuclear cells therapy could improve hip function in term of Hairrs score (MD=8.11, 95%CI 4.36 to 11.87, P<0.000 1), Merle D`Aubigné Postel score (MD=2.23, 95%CI 0.97 to 3.49, P=0.000 5), WOMAC score (MD=−10.81, 95%CI −15.80 to −5.81, P<0.000 1), Lequesne index (MD=−2.97, 95%CI −5.42 to −0.52, P=0.02) and alleviate the pain (MD=−9.13, 95%CI −12.40 to −5.86, P<0.000 01), delay the progression of radiological staging (RR=0.55, 95%CI 0.34 to 0.89, P=0.01) and reduce the rate of total hip arthroplasty (RR=0.61, 95%CI 0.43 to 0.86, P=0.005). In terms of safety, mononuclear cell therapy did not increase the rate of complications (RR=0.77, 95%CI 0.33 to 1.83, P=0.56).ConclusionsThe current evidence shows that autologous mononuclear cells therapy is a safe and effective way for osteonecrosis of the femoral head. Due to limited quality and quantity of the included studies, more high quality studies are required to verify above conclusions.
Objective To investigate the effect and safety of autologous bone marrow-mononuclear cell (BM-MNC) transplantation on ischemic limb of patients with thromboangiitis obliterans (TAO). Methods Thirteen patients with TAO underwent transplantation of autologous BM-MNC into ischemic muscles of 17 lower limbs. A series of subjective indexes (improvement of pain and cold sensation) and objective indexes including increase of ankle brachial index (ABI), transcutaneous oxygen pressure (TcPO2), and improvement of foot skin ulcer were used to evaluate the effects. Results The outcomes were evaluated after 2 months of transplantation. The pain relief and improvement of cold feeling were in 15 limbs and 16 limbs, respectively. Before transplantation and 2 months after transplantation, ABI was 0.37±0.06 and 0.50±0.17, respectively (Plt;0.05), and TcPO2 of the ischemic legs were (24.59±3.36) mm Hg (1 mm Hg=0.133 kPa) and (35.00±10.44) mm Hg, respectively (Plt;0.05). ABI increased in 9 limbs. TcPO2 elevated in 14 limbs. Skin ulcer improved in 7 limbs. Thirteen patients were followed up from 4 to 18 months (average 8 months), the patients’ symptoms improved in 13 limbs. ABI was 0.45±0.14, which wasn’t different from those before transplantation and 2 months after transplantation (Pgt;0.05). TcPO2 was (33.24±10.43) mm Hg, which was different from those before transplantation and 2 months after transplantation (Plt;0.05) and was elevated in 12 limbs. Skin ulcer healing was in 5 limbs. The ischemic symptoms in 2 patients were not relieved. There was no mortality and high level amputation. The following complications, such as proliferative retinopathy, malignant tumor, myocardial infarction, stroke or hemangioma, were not found in all patients.Conclusion In patients with TAO, intramuscular transplantation of autologous BMMNC is a safe and effective method, and may improve symptoms and accelerate the healing of skin ulcer.
The bone marrow mononuclear cell（BMMNC） subset comprises mesenchymal stem cells, hematopoietic stem cells, endothelial progenitor cells. These cells can differentiate into cardiomyocytes, vascular endothelial cells and smooth muscle cells, and they can also release a wide array of cytokines that exert their effects on surrounding cells, including inducing neovascularization, preventing apoptosis of home cells and homing of endogenous systemic repairing cells. Many trials have been developed to evaluate the effect of bone marrow mononuclear cell transplantation in treating ischemia heart diseases in this country and others. Several routes have been used to deliver these cells to human myocardium or to the coronary circulation in these trials, such as intracoronary injection, intravenous infusion, direct injection into the ventricular wall, or transepicardial/transendocardial infusions，and the cells are constructed into fragmented cell sheets to improve cell retention, or some cytokines are used to enhance therapeutic effect. Although the results of the recent clinical trials in this area are rather conflicting, these therapeutic approaches seem to be promising forthe treatment of ischemic heart disease. In this review, many aspects of bone marrow mononuclear cell transplantation in myocardial infarction are summarized such as the mechanism, delivery routes, retaining of cells, homing, survival and future development, etc.
ObjectiveTo investigate activation of phosphatidylinositol 3 hydroxykinase (PI3K)/AKT pathway and sphingosine 1-phosphate receptor 2 (S1PR2) in peripheral blood mononuclear cells (PBMCs) of acute obstructive cholangitis (AOC) rats and their effects on systemic inflammation in rats.Methods① In vitro experiment: The isolated PBMCs from the rats were divided into 4 groups: a control group, LY294002 treatment group, lipopolysaccharide (LPS) treatment group, and LPS+LY294002 treatment group. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the supernatant were detected and the phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the PBMCs were detected. ② In vivo experiment: The rats were randomly divided into four groups: a control group, LY294002 treatment group, AOC model group, and AOC+LY294002 treatment group. The survival rate of rats was recorded, the liver function (ALT, AST, and TBIL), TNF-α, and IL-6 levels in the serum were detected. The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the PBMCs of the rats were detected. Results① The results of in vitro experiment: The levels of TNF-α and IL-6 in the LPS+LY294002 treatment group were significantly lower than those in the LPS treatment group (P<0.050). The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the LPS+LY294002 treatment group were significantly lower than those in the LPS treatment group (P<0.050). ② The results of in vivo experiment: The survival rate of rats in the AOC+LY294002 treatment group was higher than those in the AOC group. The serum levels of ALT, AST, TBIL, TNF-α, and IL-6 in the AOC+LY294002 treatment group were significantly lower than those in the AOC model group (P<0.050). The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the AOC+LY294002 treatment group were significantly lower than those in the AOC model group (P<0.050).ConclusionInhibition of activation of PI3K/AKT pathway in PBMCs can inhibit expression of S1PR2, then alleviate systemic inflammatory response induced by AOC in rats.
【摘要】 目的 了解乙型肝炎病毒（HBV）X基因在HBV相关肝病肝移植受体术后外周单个核细胞（PBMC）和骨髓CD34+细胞内的整合情况及其对乙肝疫苗接种的影响。 方法 采集1999年6月-2005年11月25例HBV相关肝病肝移植受体的外周静脉血及其中23例的骨髓血，以密度梯度离心结合单克隆免疫磁珠分离法获取外周血单个核细胞及骨髓CD34+细胞后，提取细胞DNA。因HBV X基因的整合频率最高，设计HBV X基因的特异引物，进行HBV-Alu-PCR，终产物进行电泳并回收、连接载体、筛选扩增后测序，检测有无X基因整合。 结果 经PCR后电泳及测序分析，25例HBV相关肝病肝移植受体术后的PBMC内未检测出HBV X基因的整合，其中采集到骨髓标本的23例CD34+细胞中亦未检测到HBV X基因的整合。 结论 肝移植术后受体体内HBV微生态的剧烈改变，使HBV整合的基本条件丧失，在此情况下，外周免疫细胞及骨髓造血干/祖细胞不是发生HBV整合的适宜场所，乙肝疫苗接种效果与HBV X基因整合关系不明确。【Abstract】 Objective To investigate whether hepatitis B virus HBV X gene integrates in peripheral blood mononuclear cell (PBMC) and bone marrow CD34+ cells from HBV related liver disease recipients after liver transplantation. Methods Between June 1999 and November 2005, PBMC were obtained from 25 HBV related liver disease recipients after liver transplantation and bone marrow CD34+ cells obtained from 23 cases among them. The cellular DNA was extracted by DNA isolation and purification kit following the manufacture’s instructions. Specific primers to HBV X gene and to human Alu repeats were used to amplify the virus integration through a 3-round hemi-nest PCR. The PCR final product was judged by 1.2% agarose electrophoresis, ligated to T vector, proliferated in E. coil 5α and sequenced. Results According to agarose electrophoresis and sequencing analysis, there were no HBV X gene integration in PBMC and bone marrow CD34+ cells from HBV related liver transplant recipients after surgery. Conclusions Because of the radical change of HBV microecological environment in HBV related liver transplant recipients after operation, the fundamental condition of HBV integration has been lost, which led PBMC and bone marrow CD34+ cells not suit to HBV X gene integrate to human genome. And the impact of HBV X gene integration on HBV vaccination is still undefined.
Objective To explore the middle-term outcome of autologous bone marrow mononuclear cells transplantation in the treatment of lower l imb ischemia. Methods From March 2003 to June 2005, 65 patients with lower l imb ischemia were treated by autologous bone marrow mononuclear cells transplantation. Of the patients, there were 50 males and 15 females, with a mean age of 66.5 years (range 36-89 years), including 4 cases of simple arteriosclerotic occlusion,5 cases of thromboangiitis obl iterans and 56 cases diabetic lower l imb ischemia. A total of 400 mL bone-marrow blood were extracted from the posterior superior il iac crest. And then the mononuclear cells were isolated from the bone-marrow blood in the laboratory. The amount of transplantation bone marrow mononuclear cells was (0.60-1.80) × 109 (mean 1.05 × 109). Twelve patients received cell transplantation from two to four times and the other patients one time. According to the improvement of cl inical finding, the outcome was evaluated. Results All the patients were followed up for 8-56 months (mean 21.5 months). There were 8 deaths, and the mortal ity was 12.3%; 5 were due to myocardial infarction and heart failure and 3 were due to cerebral infarction. The general effective rate was 70.8% (46/65) and the recurrent rate was 10.7% (7/65). Of them, the response to treatment lasted over 12 months in 42 cases, accounting for 91.3% (42/46); over 24 months in 24 cases, accounting for 52.2% (24/46); and over 37 months in 12 cases, accounting for 26.1% (12/46). The effective rates were 100% in 12 patients who received 2-4 times transplantation and 64.2% in 53 patients who received 1 time transplantation, showing statistically significant difference between them (P lt; 0.001). Conclusion The middle-term outcome of autologous bone marrow mononuclear cells transplantation show that it is a feasible and simple method for treatment of lower l imb ischemia.