ObjectiveTo obtain a construct chimeric antigen receptor macrophages （CAR-M） targeting disialoganglioside GD2 （GD2） and harboring the intracellular domain of macrophage-specific activating signaling adaptor protein DNAX-activating protein 12 （DAP12）. MethodsA CAR molecule targeting GD2 with the DAP12 intracellular domain as the signaling module ［Anti-GD2 CAR （DAP12）］ was designed， synthesized， and cloned into an adenoviral vector. This virus was used to infect PMA-differentiated THP-1 macrophages to construct green fluorescent protein （GFP）-labeled Anti-GD2 CAR THP-1 cells. CAR expression was confirmed using live-cell imaging. In vitro co-culture assays were performed to evaluate the phagocytic function of empty， Anti-GD2 CAR-M （CD3ζ）， and Anti-GD2 CAR-M （DAP12） groups against glioma cells with high （T98G） and low （U251） GD2 expression. ResultsThe CAR adenovirus efficiently transduced THP-1 macrophages， resulting in successful generation of Anti-GD2 CAR-M. Flow cytometry analysis following co-culture with glioma cells showed that compared to the empty group， the Anti-GD2 CAR-M （CD3ζ） group exhibited significantly enhanced phagocytosis （P<0.05）. Furthermore， compared to the Anti-GD2 CAR-M （CD3ζ） group， the Anti-GD2 CAR-M （DAP12） group demonstrated a further increase in phagocytic capacity specifically against GD2-high T98G cells （P<0.05）. ConclusionAnti-GD2 CAR-M （DAP12） cells are successfully established by transducing PMA-differentiated THP-1-M with recombinant Anti-GD2 CAR （DAP12） adenovirus. The engineered CAR-M can specifically target phagocytose glioma cells with high GD2 expression.

ObjectiveTo obtain stably transfected chimeric antigen receptor macrophages （CAR-M） targeting disialosylganglioside （GD2） based on the constitutive expression of myeloid differentiation factor 88 （MYD88）. MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ （cluster of differentiation 3 zeta chain）-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages （THP-1M）， induced by phorbol 12-myristate 13-acetate （PMA） differentiation， were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein （GFP） tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M， Anti-GD2 CAR-M（CD3ζ）， and Anti-GD2 CAR-M（CD3ζ-MYD88） groups against glioma cells was evaluated in vitro through co-culture experiments， and their polarization status was also analyzed. ResultsCAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells， flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88） group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）. Compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88）CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）， and the expression of inflammatory cytokines interleukin （IL）-6 and IL-12 was increased in this group （P<0.05）. ConclusionThis study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity， demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.

Objective To analysis the effect of salbutamol combined with noninvasive ventilator on TNF-α,IL-6,IL-8 and serum CK-MB activity in patients with chronic obstructive pulmonary disease with respiratory failure.Methods 60 patients who were diagnosed with chronic obstructive pulmonary disease with respiratory failure were collected.All patients were randomly divided into experimental group and control group,30 cases in each group. Patients in the control group received noninvasive ventilator treatment, patients in the experimental group were given salbutamol treatment on the basis of control group treatment, after the treatment, the serum levels of TNF-α,IL-6,IL-8 and serum CK-MB activity were detected in all patients.Results After treatment, compared with control group, the serum level of TNFαwas lower in the experimental group,and the difference was statistically significant(P<0.05); the serum level of IL-6 and IL-8 were lower in the experimental group(P<0.05); the serum CK-MB activity was lower in the experimental group(P<0.05).Conclusion The salbutamol combined with noninvasive ventilator can significantly reduce the serum levels of TNF-α, IL-6, IL-8 and serum CK-MB activity in patients with chronic obstructive pulmonary disease with respiratory failure, reduce the inflammatory reaction and reduce the myocardial damage,and have a guiding significance for clinica.

Peripheral neuropathy induced by human immunodeficiency virus (HIV) infection and antiretroviral therapy is not only difficult to distinguish in clinical practice, but also difficult to relieve the pain symptoms by analgesics because of the severity of the disease at the later stage. Hence, to explore the mechanisms of HIV-related neuropathy and find new therapeutic options are particularly important for relieving neuropathic pain symptoms of the patients. In the present study, primary cultured embryonic rat dorsal root ganglion (DRG) neurons were used to determine the neurotoxic effects of HIV-gp120 protein and/or antiretroviral drug dideoxycytidine (ddC) and the therapeutic actions of insulin-like growth factor-1 (IGF-1) on gp120- or ddC-induced neurotoxicity. DRG neurons were exposed to gp120 (500 pmol/L), ddC (50 micromol/L), gp120 (500 pmol/L) plus ddC (50 micromol/L), gp120 (500 pmol/L) plus IGF-1 (20 nmol/L), ddC (50 micromol/L) plus IGF-1 (20 nmol/L), gp120 (500 pmol/L) plus ddC (50 micromol/L) plus IGF-1 (20 nmol/L), respectively, for 72 hours. The results showed that gp120 and/or ddC caused neurotoxicity of primary cultured DRG neurons. Interestingly, the severity of neurotoxicity induced by gp120 and ddC was different in different subpopulation of DRG neurons. gp120 mainly affected large diameter DRG neurons (>25 microm), whereas ddC mainly affected small diameter DRG neurons (< or =25 microm). IGF-1 could reverse the neurotoxicity induced by gp120 and/or ddC on small, but not large, DRG neurons. These data provide new insights in elucidating the pathogenesis of HIV infection- or antiretroviral therapy-related peripheral neuropathy and facilitating the development of novel treatment strategies.
Analgesics ; Animals ; Diagnosis-Related Groups* ; Ganglia, Spinal ; HIV ; Humans ; Insulin-Like Growth Factor I* ; Neuralgia ; Neurons* ; Peripheral Nervous System Diseases ; Rats ; Zalcitabine*

Objective To investigate the value of MSCTA in diagnosing the origin of abdominal ectopic ovarian tumor. Methods Twenty-eight patients with 30 ectopic ovarian tumors that mainly manifested as abdominal lumps were evaluated with MSCTA retrospectively. Vascular three-dimensional reconstruction by VR based on add vessel (AV) and MIP was performed for all these patients after MSCT scanning.The origin of these tumors was judged by the feeding artery and ovarian vascular pedicle (OVP) sign.The result was compared with surgical findings.Results The maximum diameter of the 30 ectopic ovarian tumors ranged from 5.2 to 19.5 cm with an average of 9.3 cm.Of the 30 ectopic ovarian tumors, the ramus ovaricus arteriae uterinae (ROAU) and ovarian artery (OA) were demonstrated in 96.7% (29/30) and 56.7% (17/30) respectively.The feeding artery of the tumor was ROAU (29 cases) or OA (1 case), and OA participated in feeding the tumor in 16 cases.The accuracy rate of diagnosis of ectopic ovarian tumors according to the feeding artery was 100% (30/30).According to OVP the accuracy rate was 85.2% (23/27) with an incidence of 76.7% (23/30).Conclusion By showing ROAU and OA as the feeding artery and ovarian vein (OV) as the draining vein, MSCTA can accurately diagnose ectopic ovary tumors in the abdomen.

Objective To investigate the expression of heat shock protein 70(HSP70) and c-fos,the presence of apoptosis and the neuroprotective effects of Cudrania tricuspidata root extrate(Ecr) after focal cerebral ischemic reperfusion in rats.Methods Models of middle cerebral artery occlusion(MCAO) were used in this study.The rats in Ecr pretreated ischemia reperfusion(IR) group were fed with Ecr(2 ml tid) for 5 days before MCAO.The brain specimen were took at different reperfusion times: 1 h,6 h,12 h,24 h,3 d and 7 d after recirculation.Then immunohistochemistry,insitu hybridization,terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) were used to detect the expression of HSP70,c-fos gene and cell apoptosis in the brains.Positive results were semiquantitatively analyzed.Results IR could induce the expression of HSP70 and c-fos.HSP70 was detected at 6 h following reperfusion and peaked at 24 h both in cortical and basal ganglia regions.c-fos was markedly expressed at 1 h and the level peaked at 6 h in the ischemic hemispheres,and then reduced gradually.The TUNEL positive cells were markedly observed at 6 h.After treatment with Ecr,the positive reaction cells both of HSP70 and c-fos were significantly increased(all(P