Fabrication of Adipose-Derived Mesenchymal Stem Cell Aggregates using Biodegradable Porous Microspheres for Injectable Adipose Tissue Regeneration

Injectable mesenchymal stem cell aggregates were formed using hyaluronic acid (HA)-immobilized porous biodegradable microspheres for adipose tissue regeneration. Adipose tissue-derived mesenchymal stem cells (AMSCs) were aggregated in a controlled manner and differentiated into adipocytes by cultivating in a stirred suspension bioreactor. The resultant cellular aggregates were approx. 1700 μm in diameter and exhibited fully differentiated adipocytes, as shown by immunocytochemistry and RT-PCR. The cultured aggregates could be smoothly injected into the subcutaneous region of mice through a syringe needle due to their soft elasticity and deformability. The in vivo regenerated adipose tissue maintained a proper dimension and shape, showing natural adipose tissue characteristics, as demonstrated by various histological staining procedures. HA-immobilized microspheres significantly enhanced cell differentiation during 3D cultivation, and tissue regeneration when implanted in vivo, compared to unmodified porous microspheres. This study showed that AMSC cellular aggregates prepared by using porous microspheres could be delivered in an injectable manner into the body and could have great therapeutic potential for soft tissue augmentation and reconstruction.     

Comparison between C57Bl/6 and C57Bl/10 mycobacterial mouse pleurisy with respect to cellular migration and nitric oxide production

Mycobacterium bovis-BCG (BCG) and Mycobacterium leprae (ML) have opposite inflammatory properties. Mycobacteria-induced pleurisy in C57Bl/6 and C57Bl/10 mice was evaluated to establish if their innate responses could be comparable, verifying cellular migration and nitrite production. Kinetic responses after ML or BCG intrathoracic injection were compared in those mice, sharing the H-2^b MHC haplotype. BCG led to acute eosinophilia and late neutrophilia in both mice. In C57Bl/6 late pleurisy, monocytes and neutrophil recruitment was dose- and iNOS-dependent, inhibited by methotrexate but not by indomethacin. Pleural macrophages released nitrites ex vivo after 7 days of BCG stimulus, without “priming” and blocked by the nitrite inhibitor L-N5-(1-iminoethyl)-ornithine (L-NIO). ML did not induce cellular migration or nitrite production, independent of the mouse strain, timing, or number of bacilli. Although these mycobacteria have high homology, there was no effect of ML on BCG-evoked secondary cellular recruitment. Both C57Black mice trigger similar onset of inflammatory responses to these mycobacteria, so far can alternatively be used in experimental studies.     

温针,穴位注射治疗类风湿性关节炎及其对细胞免疫功能的影响

本文采用温针、穴位注射治疗类风湿性关节炎５４例，分为两组并加常对照组观察治疗前后外周血Ｔ淋巴细胞亚群、ＮＫ细胞活性、白细胞介素Ⅱ（ＩＬ－２）等项免疫指标的变化。实验结果：温针与穴位注射追风速两组治疗前后外周血Ｔ淋巴细胞亚群变化不明显（Ｐ＞０．０５），ＮＫ细胞活性及ＩＬ－２治疗前均低于正常人组，经治疗后两组均有不同程度升高（Ｐ＜０．０１）。进一步证明了针灸对机体免疫功能的调节作用。     

Clearance of Senescent Cells From Injured Muscle Abrogates Heterotopic Ossification in Mouse Models of Fibrodysplasia Ossificans Progressiva

Fibrodysplasia ossificans progressiva (FOP) is a rare genetic disease caused by mutations in activin A receptor type I/activin-like kinase 2 (ACVR1/ALK2), a bone morphogenetic protein (BMP) type I receptor, resulting in the formation of extraskeletal or heterotopic ossification (HO) and other features consistent with premature aging. During the first decade of life, episodic bouts of inflammatory swellings (flare-ups) occur, which are typically triggered by soft tissue trauma. Through an endochondral process, these exacerbations ultimately result in skeletal muscles, tendons, ligaments, fascia, and aponeuroses transforming into ectopic bone, rendering movement impossible. We have previously shown that soft tissue injury causes early FOP lesions characterized by cellular hypoxia, cellular damage, and local inflammation. Here we show that muscle injury in FOP also results in senescent cell accumulation, and that senescence promotes tissue reprogramming toward a chondrogenic fate in FOP muscle but not wild-type (WT) muscle. Using a combination of senolytic drugs we show that senescent cell clearance and reduction in the senescence associated secretory phenotype (SASP) ameliorate HO in mouse models of FOP. We conclude that injury-induced senescent cell burden and the SASP contribute to FOP lesion formation and that tissue reprogramming in FOP is mediated by cellular senescence, altering myogenic cell fate toward a chondrogenic cell fate. Furthermore, pharmacological removal of senescent cells abrogates tissue reprogramming and HO formation. Here we provide proof-of-principle evidence for senolytic drugs as a future therapeutic strategy in FOP. © 2021 American Society for Bone and Mineral Research (ASBMR).     

Differential Effects of Deoxycholic Acid on Proliferation of Neoplastic and Differentiated Colonocytes in Vitro

The secondary bile acid deoxycholic acid isbelieved to be a promoter of large bowel cancer, in partby inducing colonic epithelial proliferation. Theeffects of deoxycholic acid on [³H]thymidineincorporation by the human colon cancer cell line HT29 andtwo differentiated subclones were measured and compared.The subclone HT29-C1 has features of mature absorptivecells and HT29-N2 cells secrete mucus under cholinergic control. The three cell lines were treated withdeoxycholic acid (DCA) at concentrations of 0, 5, 10,50, 100, 150, and 300 M for 3, 6, 9, 15, 24, and 48hr. A significant increase in proliferation was noted in HT29 cells only at 6 hr with 5 and 10muM deoxycholic acid. Neither the subclone HT29-C1, norHT29-N2 cells exhibited significant change in[³H]thymidine incorporation with DCA at theseconcentrations or time points. Higher doses of deoxycholicacid above 50 M and duration of exposure greaterthan 24 hr were cytotoxic to all three cell lines. Theproliferative effects of DCA in HT29 cells were not paralleled by changes in protein kinase Cactivity or protein kinase C isoform expression.Quantitative and qualitative differences in PKC isoformexpression were not noted in the three cell lines used in this study. The proliferative effects of DCAon HT29 cells appear to be independent of the PKC signaltransduction pathway.     

五子衍宗丸对小鼠免疫功能的影响

本文探讨了五子衍宗丸对小鼠免疫功能的影响。结果表明,五子衍宗丸对正常小鼠外周血白细胞总数及酸性非特异性脂酶阳性淋巴细胞百分率无明显影响(P>0.05),对正常小鼠脾细胞产生抗体功能亦无明显影响(P>0.05),但对正常小鼠及免疫受抑小鼠的单核吞噬细胞功能有明显的激活增强作用(P<0.01),对免疫受抑小鼠外周血白细胞总数及酸性非特异性脂酶阳性淋巴细胞百分率的减少有防治作用(P<0.01),对免疫受抑小鼠脾细胞产生抗体功能有明显的促进作用。结果提示,五子衍宗丸对正常小鼠的部分非特异性免疫功能有一定的增强作用,而对免疫受抑小鼠的非特异性免疫功能及特异性免疫功能(包括细胞免疫和体液丸疫)均有明显的增强作用。     

肝癌患者集束电极射频消融治疗前后T淋巴细胞及红细胞免疫功能的研究

目的　研究肝癌患者接受射频消融治疗前后机体T淋巴细胞及红细胞免疫功能的变化。方法　分析 12 0例肝细胞肝癌患者射频消融治疗前和治疗后 3d ,7d和 14d外周血淋巴细胞亚群(T 3 ,T 4,T 8,T4/T 8)和红细胞C 3b受体花环 (RBC C3bR )及免疫复合物花环 (RBC ICR )形成率。结果　所有肝癌患者在射频消融治疗后 7d和 14d ,T 3细胞 ,T 4细胞 ,T 4/T 8及RBC C 3bR ,RBC ICR形成率均明显高于治疗前 (P <0 .0 5 )。结论　采用射频消融治疗肝癌 ,可以改善患者的细胞免疫状态。     

柴葛颗粒剂的降温作用与免疫调节功能

为了探讨由柴胡、葛根、麻黄、杏仁、石膏、甘草等组成的 ,用于治疗上感高热之证 ,具有卫气两清作用的中药复方柴葛颗粒的降温及调节免疫作用 ,采用腹腔巨噬细胞功能测定、免疫功能测定等方法观察了柴葛颗粒对伤寒、副伤寒甲、乙三联菌苗及内毒素诱发家兔体温升高的调节作用。结果表明 ,在三联菌苗试验中 ,柴葛颗粒 6.0g kg剂量组解热作用优于银黄口服液组 (P <0 .0 1) ,在内毒素致热实验中 ,以4.0g kg剂量组的效果最好。柴葛颗粒 3个剂量组 ( 4.0g kg、 2 .0g kg、 1.0g kg)均能增加小鼠吞噬细胞的吞噬百分率和吞噬指数 ,并呈量效关系。恶葛颗粒能增加小鼠脾重、E花结数 ,能增加小鼠胸腺质量 ,对氢化泼尼松 (PDS)免疫小鼠的脾及胸腺萎缩有一定的对抗作用 ,柴葛颗粒不能提高正常小鼠及PDS免疫抑制小鼠的空斑形成细胞 (PEC)值和IgG、IgM含量。可见柴葛颗粒具有解热作用 ,能提高动物T细胞介导的细胞免疫功能 ,对抗体分泌细胞及抗体水平影响不大     

Involvement of Rap-1 activation and early termination of immune synapse in CTLA-4-mediated negative signal

Abstract

Cytotoxic T lymphocyte antigen 4 (CTLA-4) is a T cell co-stimulation receptor that delivers inhibitory signals upon activation. This inhibitory effect by CTLA-4 requires activation of small GTPase Rap-1. However, the precise mechanism underlying these negative signals remains unclear. Here, we show that CTLA-4-induced suppression of IL-2 production correlates with rapid destabilization of immunological synapse (IS) formation in murine normal T cell clones. Overexpression of Spa-1, a Rap-1-specific GTPase activating protein (GAP), abolished both Rap-1 activation and IL-2 suppression induced by CTLA-4. Although we failed to find any specific inhibition of activation of early signals upon CTLA-4 engagement, we found that CTLA-4 specifically up-regulates cell motility and suppresses prolonged accumulation of Talin at the contact area with antigen presenting cells upon antigen stimulation. These results suggest that Rap-1 is activated upon CTLA-4 ligation and mediates inhibitory signals through prevention of IS formation.