Objective: To investigate the effect of the AML1-ETO (AE) fusion gene on the biological function of U937 leukemia cells by establishing a leukemia cell model that induces AE fusion gene expression. Methods: The doxycycline (Dox) -dependent expression of the AE fusion gene in the U937 cell line (U937-AE) were established using a lentivirus vector system. The Cell Counting Kit 8 methods, including the PI and sidanilide induction, were used to detect cell proliferation, cell cycle-induced differentiation assays, respectively. The effect of the AE fusion gene on the biological function of U937-AE cells was preliminarily explored using transcriptome sequencing and metabonomic sequencing. Results: ①The Dox-dependent Tet-on regulatory system was successfully constructed to regulate the stable AE fusion gene expression in U937-AE cells. ②Cell proliferation slowed down and the cell proliferation rate with AE expression (3.47±0.07) was lower than AE non-expression (3.86 ± 0.05) after inducing the AE fusion gene expression for 24 h (P<0.05). The proportion of cells in the G(0)/G(1) phase in the cell cycle increased, with AE expression [ (63.45±3.10) %) ] was higher than AE non-expression [ (41.36± 9.56) %] (P<0.05). The proportion of cells expressing CD13 and CD14 decreased with the expression of AE. The AE negative group is significantly higher than the AE positive group (P<0.05). ③The enrichment analysis of the transcriptome sequencing gene set revealed significantly enriched quiescence, nuclear factor kappa-light-chain-enhancer of activated B cells, interferon-α/γ, and other inflammatory response and immune regulation signals after AE expression. ④Disorder of fatty acid metabolism of U937-AE cells occurred under the influence of AE. The concentration of the medium and short-chain fatty acid acylcarnitine metabolites decreased in cells with AE expressing, propionyl L-carnitine, wherein those with AE expression (0.46±0.13) were lower than those with AE non-expression (1.00±0.27) (P<0.05). The metabolite concentration of some long-chain fatty acid acylcarnitine increased in cells with AE expressing tetradecanoyl carnitine, wherein those with AE expression (1.26±0.01) were higher than those with AE non-expression (1.00±0.05) (P<0.05) . Conclusion: This study successfully established a leukemia cell model that can induce AE expression. The AE expression blocked the cell cycle and inhibited cell differentiation. The gene sets related to the inflammatory reactions was significantly enriched in U937-AE cells that express AE, and fatty acid metabolism was disordered.目的： 通过建立可诱导表达AML1-ETO（AE）融合基因的白血病细胞模型，研究AE融合基因对U937白血病细胞生物学功能的影响。 方法： 利用慢病毒载体系统，构建强力霉素（Dox）依赖的可诱导表达AE融合基因的U937细胞系（U937-AE）。在AE融合基因表达前后，分别采用CCK-8法、流式细胞术进行细胞增殖、周期、诱导分化检测，并进行转录组学测序和代谢组学测序分析，初步探讨AE融合基因对白血病细胞生物学功能的影响。 结果： ①成功构建Dox依赖的Tet-on调控系统，调控AE融合基因在U937-AE细胞中稳定表达。②诱导AE融合基因表达24 h后，表达AE的U937-AE细胞增殖倍率为3.47±0.07，低于AE阴性组的3.86±0.05（P<0.05）；处于G(0)/G(1)期的细胞比例为（63.45±3.10）%，明显高于AE阴性组的（41.36±9.56）%（P<0.05）；表达CD13、CD14的细胞比例较AE阴性组明显下降（P<0.05）。③转录组学测序进行基因集富集分析显示，与静息相关、NF-κB和干扰素α/γ应答相关的炎症反应和免疫调节的基因集被明显富集在表达AE的U937-AE细胞。④U937-AE细胞表达AE融合基因后脂肪酸代谢发生紊乱，AE阳性组细胞的部分中、短链脂肪酸酰基肉碱的代谢物浓度降低[丙酰基-L-肉碱：AE阳性组0.46±0.13，AE阴性组1.00±0.27（P<0.05）]；部分长链脂肪酸酰基肉碱的代谢物浓度升高[十四烷酰肉碱：AE阳性组1.26±0.01，AE阴性组1.00±0.05（P<0.05）]。 结论： 成功建立可诱导表达AE融合基因的白血病细胞模型。AE融合基因表达使U937-AE细胞增殖变慢、周期阻滞、分化受抑，炎症反应和免疫调节的相关基因集被明显富集，细胞的脂肪酸代谢发生紊乱。.
Abstract Based on daily data from Japanese 55‐year Reanalysis (JRA‐55), this study examines the formation and maintenance mechanisms of the subseasonal eastern Pacific (EP) pattern from an energetics perspective. Results show that the baroclinic energy conversion (CP B ) acts as an important energy source to overcome the thermal damping caused by the transient eddies and as a major kinetic energy (KE) source. The feedback forcing of transient eddies (CK E ) acts as a major KE source during the growing stage and a major KE sink during the decay of the EP pattern. At the same time, the barotropic energy conversion (CK B ) also makes substantial positive KE contribution. The results also suggest that the EP dynamics differ among different tropospheric layers. In the upper layer, CP B , CK B , and the anomalous geopotential height flux divergence (AGHFD) term all act as major KE source, while CK E acts as a major KE source (sink) during the growth (decay) stage of the EP pattern. The middle layer is mainly driven by CP B and AGHFD: CP B acts as a KE source while AGHFD as a sink. In the lower layer, CP B and AGHFD act as major KE sources with the later larger than the former, while the friction term damps the EP pattern heavily. It is found that the vertical component of the divergence of geopotential height flux anomaly plays a crucial role in the dynamic coupling of the different vertical parts of the EP pattern.
Objective:To summarize the clinical features of lumbar disc herniation and treatment points.Methods:We retrospectively analysed clinical data of 260 patients with lumbar disc herniation.Results:of straight leg raising test results and imaging was 100%, the treatment rate of 88.08%good,efficient 100%.Conclusion:The waist,leg and hip pain,numbness,body position changes,movement disorders,sensory disturbances,muscle atrophy of lumbar disc herniation are all the major clinical manifestations;straight leg raising test with high test can be used as important reference for the early diagnosis,to signs according to patient,disease and other specific conditions to choose the best treatment method.
Obesity is an epidemic chronic disease which is a risk factor for a number of serious medical conditions. Intragastric balloon, as an alternative, non-surgical treatment approach for the management of obesity, was rejected in previous evidence-based reviews. The object of this review is updating the data and confirming the safety and efficacy of the procedure. In this review, systematic literature retrieve of MEDLINE, EMBASE, CENTRAL and other information sources was performed from inception to December 2014. The quality of selected studies was assessed and meta-analyses of weighted mean differences were made using the inverse variance method. Meta-analyses presented significant effect sizes of -8.9 kg, -3.1 kg/m(2) and -21.0% for SMG as well as of -1.5 kg and -1.2 kg/m(2) for LSG, favoring the intervention group. Safety analysis showed that minor complications occurred at a high rate in intervention group, however, no serious or fatal complication was reported in these studies. In conclusion, the current review presents that short-term efficacy for 6 months treatment of intragastric balloon in association with conservative therapy is clinically significant.
Introduction Metastatic spinal cancer is a common condition that may lead to spinal instability, pain and paralysis. In the 1980s, surgery was discouraged because results showed worse neurological outcomes and pain compared with radiotherapy alone. However, with the advent of modern imaging and spinal stabilisation techniques, the role of surgery has regained centre stage, though few studies have assessed quality of life and functional outcomes after surgery. Objective We investigated whether surgery provides sustained improvement in quality of life and pain relief for patients with symptomatic spinal metastases by analysing the largest reported surgical series of patients with epidural spinal metastases. Methods A prospective cohort study of 922 consecutive patients with spinal metastases who underwent surgery, from the Global Spine Tumour Study Group database. Pre- and post-operative EQ-5D quality of life, visual analogue pain score, Karnofsky physical functioning score, complication rates and survival were recorded. Results Quality of life (EQ-5D), VAS pain score and Karnofsky physical functioning score improved rapidly after surgery and these improvements were sustained in those patients who survived up to 2 years after surgery. In specialised spine centres, the technical intra-operative complication rate of surgery was low, however almost a quarter of patients experienced post-operative systemic adverse events. Conclusion Surgical treatment for spinal metastases produces rapid pain relief, maintains ambulation and improves good quality of life. However, as a group, patients with cancer are vulnerable to post-operative systemic complications, hence the importance of appropriate patient selection.
Objective: This study aimed to create a type of CAR-T cells that targets LMP1 antigen and study its immunotherapeutic effect on LMP1-positive hematological malignancies. Methods: To generate LMP1 CAR-T cells, a plasmid expressing LMP1 CAR was created using molecular cloning technology, and T cells were infected with LMP1 CAR lentivirus. The effects of LMP1 CAR-T cells on specific cytotoxicity against LMP1-positive tumor cell lines infected with the EB virus had been confirmed. Results: ① LMP1 protein expressing on EB virus-positive lymphoma cells surface was verified. ② The LMP1 CAR-expressing plasmid was created, and LMP1 CAR-T cells were obtained by infecting T cells with a lentivirus packaging system, with an infection efficiency of more than 80% . ③LMP1 CAR-T cells have a 4∶1 effect-to-target ratio in killing LMP1-positive lymphoma cells. The killing effect of LMP1 CAR-T cells on Raji cells was enhanced after 48 h of coculture, but there was no significant killing effect on Ramos, which are LMP1-negative lymphoma cells. ④After coculture with LMP1-positive lymphoma cells at a ratio of 1∶1 for 5 h, the degranulation effect was enhanced. The proportion of CD107a(+) T cells in the LMP1 CAR-T cell treatment group was significantly higher than that in the vector-T cell group [ (13.25±2.94) % vs (1.55±0.05) % , t=3.972, P=0.017]. ⑤After coculture with LMP1-positive lymphoma cells, the proportion of CD69(+) and CD25(+) T cells in the LMP1 CAR-T cell group was significantly higher than that in vector-T cell group [ (7.40±0.41) % vs (3.48±0.47) % , t=6.268, P=0.003; (73.00±4.73) % vs (57.67±2.60) % , t=2.842, P=0.047]. ⑥After coculture with LMP1-positive lymphoma cells, cytokine secretion in the LMP1 CAR-T cell group was higher than that in the vector-T cell group [interferon-gamma: (703±73) ng/L vs (422±87) ng/L, t=2.478, P=0.068; tumor necrosis factor-alpha: (215±35) ng/L vs (125±2) ng/L, t=2.536, P=0.064]. Conclusion: In this study, we found that the LMP1 protein is only found on the surface of the EBV-positive tumor cell. Simultaneously, we created an LMP1 CAR-expressing plasmid and obtained LMP1 CAR-T cells by infecting T cells with a lentivirus packaging system. Furthermore, we demonstrated that LMP1 CAR-T cells could specifically kill LMP1-positive tumor cells in vitro. The degranulation and activation effects of LMP1 CAR-T cells were enhanced after coculture with LMP1-positive tumor cells, indicating a potential clinical application.目的： 制备一种靶向LMP1抗原的嵌合抗原受体T细胞（CAR-T细胞），研究其对EB病毒（EBV）阳性淋巴瘤的免疫治疗作用。 方法： 应用分子克隆技术构建二代LMP1 CAR表达质粒，通过慢病毒包装体系包装病毒后感染人T细胞，获得LMP1 CAR-T细胞，体外实验验证LMP1 CAR-T细胞对感染EBV后的LMP1阳性淋巴瘤细胞的特异性细胞毒性作用。 结果： ①LMP1蛋白表达于EBV阳性的淋巴瘤细胞表面；②成功构建了二代LMP1 CAR慢病毒载体，感染人T细胞，获取LMP1 CAR-T细胞，感染效率大于80%；③LMP1 CAR-T细胞可特异性杀伤LMP1阳性淋巴瘤细胞，当效靶比按4∶1共培养48 h后，LMP1 CAR-T细胞对Raji细胞的杀伤作用增强，对Ramos细胞无明显杀伤作用；④与LMP1阳性淋巴瘤细胞按1∶1共培养5 h后，LMP1 CAR-T细胞处理组CD107a(+) T细胞比例显著高于Vector-T细胞组[（13.25±2.94）%对（1.55±0.05）%，t＝3.972，P＝0.017]，脱颗粒效果增强；⑤与LMP1阳性淋巴瘤细胞共培养后，LMP1 CAR-T细胞组CD69(+)、CD25(+) T细胞比例显高于Vector-T细胞组[（7.40±0.41）%对（3.48±0.47）%，t＝6.268，P＝0.003；（73.00±4.73）%对（57.67±2.60）%，t＝2.842，P＝0.047]，活化效应增强。⑥与LMP1阳性淋巴瘤细胞共培养后，LMP1 CAR-T细胞组细胞因子分泌增强，高于Vector-T细胞组[IFN-γ：（703±73）ng/L对（422±87）ng/L，t＝2.478，P＝0.068；TNF-α：（215±35）ng/L对（125±2）ng/L，t＝2.536，P＝0.064]。 结论： 该研究证实EBV阳性淋巴瘤细胞表面可特异表达LMP1蛋白，成功构建了LMP1 CAR慢病毒载体并感染人T细胞，成功获得LMP1 CAR-T细胞。体外实验证实：与LMP1阳性淋巴瘤细胞共培养后，LMP1 CAR-T细胞脱颗粒效果增强，活化效应增强，高效分泌细胞因子，可特异杀伤LMP1阳性淋巴瘤细胞，具有潜在的临床应用前景。.
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