Distribution of Yeast Cell Wall Lytic Activity among Microorganisms

An α-glucosidase has been isolated from the mycelia of Penicillium purpurogenum in electrophoretically homogeneous form, and its properties have been investigated. The enzyme had a molecular weight of 120,000 and an isoelectric point of pH 3.2. The enzyme had a pH optimum at 3.0 to 5.0 with maltose as substrate. The enzyme hydrolyzed not only maltose but also amylose, amylopectin, glycogen, and soluble starch, and glucose was the sole product from these substrates. The Km value for maltose was 6.94×10^?4 m. The enzyme hydrolyzed phenyl α-maltoside to glucose and phenyl α-glucoside. The enzyme had α-glucosyltransferase activity, the main transfer product from maltose being maltotriose. The enzyme also catalyzed the transfer of α-glucosyl residue from maltose to riboflavin.     

Developing lisocabtagene maraleucel chimeric antigen receptor T-cell manufacturing for improved process,product quality and consistency across CD19+ hematologic indications

Background aimsAutologous chimeric antigen receptor (CAR) T-cell therapies have demonstrated substantial clinical benefit across several hematologic malignancies. However, patient-to-patient variability and heterogeneity of starting cellular material across patient populations and disease indications pose challenges to manufacturing consistency. Lisocabtagene maraleucel (liso-cel) is an autologous, CD19-directed, defined-composition, 4-1BB CAR T-cell product administered at equal target doses of CD8⁺ and CD4⁺ CAR⁺ T cells. Here the authors describe the optimization of the liso-cel manufacturing platform for product quality and consistency.MethodsLeukapheresis starting materials were collected from patients with large B-cell lymphoma, mantle cell lymphoma or chronic lymphocytic leukemia treated with liso-cel in clinical trials (NCT02631044 and NCT03331198). The liso-cel manufacturing process involves selection of CD8⁺ and CD4⁺ T cells from leukapheresis material followed by independent CD8⁺ and CD4⁺ T-cell activation, transduction, expansion, formulation and cryopreservation. Multivariate design of experimental approaches was utilized to optimize process conditions at both specific unit operations and across the process. Flow cytometry methods were used to assess cellular composition, memory phenotypes and cell proliferation. Antigen-specific functions, including cytokine secretion, cytolytic activity and proliferation, were assessed using endpoint assays after independent stimulation of CD8⁺ and CD4⁺ CAR⁺ T-cell product components.ResultsReductions in process duration time, optimization of drug product container and formulation and activation signal optimization led to significantly increased CAR⁺ T-cell product viability. The heterogeneity of patient-derived starting material, including low absolute lymphocyte counts in some samples, was reduced through early T-cell purification, leading to median T-cell frequencies >95% in selected materials across disease indications and limited non-T-cell impurities. These changes further increased lineage purity in CD8⁺ and CD4⁺ CAR⁺ T-cell drug products. CD8⁺ and CD4⁺ CAR⁺ T-cell component lot functional profiles demonstrated multifunctional mechanisms of action, including differential cytokine release, differential cytolytic kinetics and high frequencies of proliferating cells. Correlative analyses demonstrated strong underlying associations between starting material attributes and final CAR⁺ T-cell product phenotype.ConclusionsDespite substantial heterogeneity of starting leukapheresis material quality/composition between individual patients and across disease indications/histologies, the liso-cel manufacturing platform is robust and capable of generating a consistent drug product from diverse starting materials with a single manufacturing platform.     

Interleukin 7 Up-regulates CD95 Protein on CD4+ T Cells by Affecting mRNA Alternative Splicing: PRIMING FOR A SYNERGISTIC EFFECT ON HIV-1 RESERVOIR MAINTENANCE*

Interleukin-7 (IL-7) has been used as an immunoregulatory and latency-reversing agent in human immunodeficiency virus type 1 (HIV-1) infection. Although IL-7 can restore circulating CD4⁺ T cell counts in HIV-1-infected patients, the anti-apoptotic and proliferative effects of IL-7 appear to benefit survival and expansion of HIV-1-latently infected memory CD4⁺ T lymphocytes. IL-7 has been shown to elevate CD95 on CD4⁺ T cells in HIV-1-infected individuals and prime CD4⁺ T lymphocytes to CD95-mediated proliferative or apoptotic signals. Here we observed that through increasing microRNA-124, IL-7 down-regulates the splicing regulator polypyrimidine tract binding protein (PTB), leading to inclusion of the transmembrane domain-encoding exon 6 of CD95 mRNA and, subsequently, elevation of CD95 on memory CD4⁺ T cells. Moreover, IL-7 up-regulates cellular FLICE-like inhibitory protein (c-FLIP) and stimulates c-Jun N-terminal kinase (JNK) phosphorylation, which switches CD95 signaling to survival mode in memory CD4⁺ T lymphocytes. As a result, co-stimulation through IL-7/IL-7R and FasL/CD95 signal pathways augments IL-7-mediated survival and expansion of HIV-1-latently infected memory CD4⁺ T lymphocytes. Collectively, we have demonstrated a novel mechanism for IL-7-mediated maintenance of HIV-1 reservoir.     

Synthesis, characterization and potential application of monoacyl-cyclodextrins

Although the preparation of cyclodextrin (CD) monoesters with a variety of carboxylic acids has been already described in the literature, the direct regioselective CD acylation has proved to be critical, often requiring to be replaced with a more elaborate synthetic process. In this paper we describe the one-step preparation of several monoacylated CDs from acyclic or aromatic carboxylic acid derivatives. The ability of β-CD to enclose cupric ions in a sandwich-type manner was exploited to lead to high regioselectivity in the acylation of β-CD with benzoyl chloride, cinnamoyl chloride and phenyl acetyl chloride in water. Long chain aliphatic monoesters of α-, β- and γ-CD were best prepared in DMF. The results of our study showed that solvent and general conditions determined an overwhelming regioselectivity of acylation. ¹H, ¹³C and 2D NMR experiments could easily discriminate the position of the ester. Monoacylated CDs were evaluated as a carrier of silibinin, the inclusion complexes were prepared and characterized by thermal analysis.     

Ginsenoside Rg1 attenuates concanavalin A-induced hepatitis in mice through inhibition of cytokine secretion and lymphocyte infiltration

The effect of ginsenoside Rg1 (Rg1) on hepatic damage caused by concanavalin A (Con A) has not been fully elucidated. This study was designed to evaluate the protective effect of Rg1 on Con A-induced hepatitis in mice and explore the potential mechanisms of this effect. C57BL/6 mice were divided randomly into the following four experimental groups: phosphate-buffered saline group, Rg1 group, Con A group, Con A + Rg1 group. Mice received Rg1 (20 mg/kg) 3 h before intravenous administration of Con A (15 mg/kg). Levels of alanine transaminase, aspartate transaminase and cytokine production were measured, the amount of phosphorylated IκBα and p65 were tested, the numbers of CD4⁺ and CD8⁺ T lymphocytes infiltrated in the blood, spleen and liver were calculated, intercellular adhesion molecule-1 (ICAM-1) and interferon-inducible chemokine-10 (CXCL-10) levels were measured and histological examination of the livers was conducted. Pretreatment with Rg1 markedly reduced the elevated levels of serum aminotransferase, ameliorated liver damage and suppressed proinflammatory cytokines secretion via inhibition NF-κB activity following Con A injection of mice. Furthermore, Rg1 administration reduced ICAM-1 and CXCL-10 mRNA expression in the liver as well as the number of CD4⁺ and CD8⁺ T lymphocytes infiltrating in the liver. Rg1 reduced the incidence of liver damage through inhibition of the proinflammatory response and suppressed the recruitment of CD4⁺ and CD8⁺ T lymphocytes to the liver. These data indicate that Rg1 represents a novel agent for the treatment of T lymphocyte-dependent liver injury.     

无充气腋窝入路腔镜下单侧甲状腺癌根治术与开放性手术的疗效比较及对免疫功能和颈部功能的影响

摘要 目的：比较无充气腋窝入路腔镜下单侧甲状腺癌根治术与开放性手术的疗效及对免疫功能和颈部功能的影响。方法：回顾性分析我院2020年8月～2021年8月期间收治的行单侧甲状腺癌根治术患者80例的临床资料。根据手术方式的不同将患者分为开放组（开放性手术）和腔镜组（无充气腋窝入路腔镜下单侧甲状腺癌根治术）,例数分别为37例和43例。对比两组疗效、免疫功能、颈部功能、美容学满意度和并发症情况。结果：与开放组相比,腔镜组术中出血量更少,手术时间、住院时间更长,引流液总量更多（P<0.05）,两组中央组淋巴结清扫数组间对比无统计学差异（P>0.05）。两组CD3⁺、CD4⁺、CD4⁺/CD8⁺下降,但腔镜组高于开放组;CD8+升高,但腔镜组低于开放组（P<0.05）。两组术后3d视觉疼痛模拟评分法（VAS）评分、颈部损伤指数对比,差异无统计学意义（P>0.05）。腔镜组吞咽障碍指数低于开放组（P<0.05）。腔镜组的总满意率高于开放组（P<0.05）。两组并发症发生率组间对比无统计学差异（P>0.05）。结论：与开放性手术治疗单侧甲状腺癌相比,无充气腋窝入路腔镜下单侧甲状腺癌根治术的手术时间和住院时间虽然延长,但其对患者免疫功能影响更轻,同时还可减轻患者吞咽障碍,获得更好的美容学满意度。     

Mitochondrial mass governs the extent of human T cell senescence

The susceptibility of human CD4⁺ and CD8⁺ T cells to senesce differs, with CD8⁺ T cells acquiring an immunosenescent phenotype faster than the CD4⁺ T cell compartment. We show here that it is the inherent difference in mitochondrial content that drives this phenotype, with senescent human CD4⁺ T cells displaying a higher mitochondrial mass. The loss of mitochondria in the senescent human CD8⁺ T cells has knock‐on consequences for nutrient usage, metabolism and function. Senescent CD4⁺ T cells uptake more lipid and glucose than their CD8⁺ counterparts, leading to a greater metabolic versatility engaging either an oxidative or a glycolytic metabolism. The enhanced metabolic advantage of senescent CD4⁺ T cells allows for more proliferation and migration than observed in the senescent CD8⁺ subset. Mitochondrial dysfunction has been linked to both cellular senescence and aging; however, it is still unclear whether mitochondria play a causal role in senescence. Our data show that reducing mitochondrial function in human CD4⁺ T cells, through the addition of low‐dose rotenone, causes the generation of a CD4⁺ T cell with a CD8⁺‐like phenotype. Therefore, we wish to propose that it is the inherent metabolic stability that governs the susceptibility to an immunosenescent phenotype.     

DCs-CIK细胞免疫治疗联合化疗对转移性前列腺癌患者免疫功能及生活质量的影响

目的:探讨DCs-CIK细胞免疫(Dendritic cells-Cytokine induced killer cells,DCs-CIK)治疗联合化疗对转移性前列腺癌患者免疫功能及生活质量的影响。方法:选择106例确诊转移性前列腺癌患者随机分为观察组与对照组,每组各53例,对照组采用多西他赛联合表阿霉素~+泼尼松化疗方式进行治疗,观察组在此基础上给予DC-CIK细胞免疫治疗。比较两组患者治疗前后外周血CD3~+、CD3~+CD4~+、CD3~+CD8~+、CD3-CD56~+、CD4~+CD8~+、自然杀伤细胞(NK)、自然杀伤T细胞(NKT)表达水平;使用QLQ-C30问卷评价患者治疗前后生活质量的变化情况,观察并比较患者治疗过程中发生的不良反应情况。结果:组观察组患者治疗总有效率达73.58%远高于对照组41.51%水平(p0.05);观察组患者治疗后外周血CD3~+、CD3~+CD4~+、CD3~+CD8~+、CD3-CD56~+、CD4~+CD8~+、NK、NKT表达水平明显好于对照组(p0.05);观察组患者躯体功能、角色功能、情绪功能、认知功能、社会功能各项指标得分明显好于对照组(p0.05);两组患者接受治疗后,均有部分患者出现恶心呕吐、脱发、白细胞减少、血小板减少或肝功受损,其中观察组患者出现恶心呕吐、白细胞减少的人数明显少于对照组(p0.05)。结论:DCs-CIK细胞免疫治疗联合化疗有助于转移性前列腺癌患者的治疗,在明显改善患者免疫能力的同时有效改善患者生活质量,具有重要的临床指导意义。     

Synthesis and conformational study of peptides possessing helically arranged ΔZPhe side chains

Z-Dehydrophenylalanine (Δ^zPhe) possessing four oligopeptides, Boc-(L -Ala-Δ^zPhe-Aib)_n-OCH₃ (n = 1–4: Boc, t-butoxycarbonyl; Aib, α-aminoisobutyric acid), were synthesized, and their solution conformations were investigated by ¹H-nmr, ir, uv, and CD spectroscopy and theoretical CD calculation. ¹H-nmr (the solvent accessibility of NH groups) and ir studies indicated that all the NH groups except for those belonging to the N-terminal L -Ala-Δ^zPhe moiety participate in intramolecular hydrogen bonding in chloroform. This suggests that the peptides n = 2–4 have a 4 → 1 hydrogen-bonding pattern characteristic of 3₁₀-helical structures. The uv spectra of all these peptides recorded in chloroform and in trimethyl phosphate showed an intense maximum around 276 nm assigned to the Δ^zPhe chromophores. The corresponding CD spectra of the peptides n = 2–4 showed exciton couplets with a negative peak at longer wavelengths, whereas that of the peptide n = 1 showed only weak signals. Theoretical CD spectra were calculated for the peptides n = 2–4 of several helical conformations, on the basis of exciton chirality method. This calculation indicated that the three peptides form a helical conformation deviating from the perfect 3₁₀-helix that contains three residues per turn, and that their side chains of Δ^z Phe residues are arranged regularly along the helix. The center-to-center distance between the nearest phenyl pair(s) was estimated to be ～ 5.5 Å. The chemical shifts of the Δ^zPhe side-chain protons (H^β and aromatic H) for the peptides n = 2–4 indicated anisotropic shielding effect of neighboring phenyl group(s); the effect also supports a regular arrangement of the Δ^z Phe side chains along the helical axis. © 1993 John Wiley & Sons, Inc.     

Skin Allografting Activates Anti-tumor Immunity and Suppresses Growth of Colon Cancer in Mice

INTRODUCTION: The tumor cells could escape from the immune elimination through the immunoediting mechanisms including the generation of immunosuppressive or immunoregulative cells. By contrast, allograft transplantation could activate the immune system and induce a strong allogenic response. The aim of this study was to investigate the efficacy of allogenic skin transplantation in the inhibition of tumor growth through the activation of allogenic immune response. METHODS: Full-thickness skin transplantation was performed from C57BL/6 (H-2^b) donors to BALB/c (H-2^d) recipients that were receiving subcutaneous injection of isogenic CT26 colon cancer cells (2?×?10⁶ cells) at the same time. The tumor size and pathological changes, cell populations and cytokine profiles were evaluated at day 14 post-transplantation. RESULTS: The results showed that as compared to non-transplant group, the allogenic immune response in the skin-grafting group inhibited the growth of tumors, which was significantly associated with increased numbers of intra-tumor infiltrating lymphocytes, increased populations of CD11c⁺MHC-classII⁺CD86⁺ DCs, CD3⁺CD4⁺ T cells, CD3⁺CD8⁺ T cells, and CD19⁺ B cells, as well as decreased percentage of CD4⁺CD25⁺Foxp3⁺ T cells in the spleens. In addition, the levels of serum IgM and IgG, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were significantly higher within the tumor in skin transplant groups than that in non-transplant group. CONCLUSIONS: Allogenic skin transplantation suppresses the tumor growth through activating the allogenic immune response, and it may provide a new immunotherapy option for the clinical refractory tumor treatment.     

The in vitro effect of methylprednisolone on the processes of activation of CD4<Superscript>+</Superscript>CD45RO<Superscript>+</Superscript> T-cells from healthy subjects and rheumatoid-arthritis patients

Flow cytometry has been used to analyze the changes in the number of CD4⁺ cells that expressed surface markers of activation (CD25, CD71, HLA-DR, and CD95) in cultures of TCR-stimulated CD3⁺CD45RO⁺ Т-lymphocytes after in vivo exposure to different concentrations of methylprednisolone (MP). T-cells were obtained from healthy donors and rheumatoid arthritis (RA) patients. Suppressive action of МР on the expression of activation and proliferation markers (CD25 and CD71, respectively) by CD4⁺ T-cells was observed in all study subjects. МР increased the number of CD4⁺ HLA-DR⁺/CD95⁺ cells among the СD3⁺CD45RO⁺ cells obtained from RA patients and subjected to TCR activation, whereas the number of such cells in the control group decreased after MP treatment. The MP-induced changes in the cells subjected to TCR activation can be indicative of relative resistance of the CD4⁺CD45RO⁺HLA-DR⁺/CD95⁺ cell population in RA patients to the action of glucocorticoids and the possible role of this subpopulation in RA pathogenesis.     

Fragmentation of Penicillin-binding Protein-1Bs of Escherichia coli by Proteolytic Enzymes: A Preliminary Study on the Location of the Penicillin-binding Site on the Bifunctional Peptidoglycan Synthetase Protein

The absolute configurations of fenvalerate and other related cyanohydrin esters were studied by circular dichroism (CD) measurements and by high performance liquid chromatography (HPLC). Fenvalerate has UV absorption peaks around 278 nm associated with the ¹L_b phenyl transitions and corresponding positive CD peaks were observed around 281 nm for the enantiomers of (S)-configuration at the cyanohydrin chiral center. Most of the other cyanohydrin esters also gave positive CD peaks for the enantiomers of (S)-configuration. CD spectra in the 180 to 250 nm range were also studied.

By HPLC, the elution order of the diastereoisomers of cyanohydrin esters were closely correlated with their absolute configuration and the (RS,SR)-pair consistently eluted earlier than the (RR,SS)-pair for α-substituted phenylacetic acid esters.     

The clinical effect of Prednisone in combination with Mycophenolate mofetil on idiopathic thrombocytopenic purpura (ITP) and its influence on the level of peripheral blood T lymphocytes and NK lymphocytes

ObjectiveTo explore he curative effect and safety of Prednisone in combination with Mycophenolate in treating ITP and its influence on the level of peripheral blood T lymphocytes and NK lymphocytes.Method93 cases of ITP patients were divided into the observation group and the control group by the Random Number Table method, 48 cases for the observation group, 45 for another. Patients in the control group orally took 0.5 mg/kg Prednisone Acetate tablets daily, two times each in the morning and evening. And the observation group, based on the treatment of the control group, orally took Mycophenolate Mofetil Dispersible tablets twice a day, 1 g each time. According to patients’ conditions, 3 to 5 courses were set for treatment with 3 weeks a course. Compared PLT amount and the changing situation of inflammatory factors, CD3⁺ and CD3⁺CD95L⁺ before and after the treatment, the level of CD3⁺Caspase-3⁺ and CD3⁺Caspase-8⁺, NK⁺, NK⁺ CD95L⁺, NK⁺Caspase-3⁺, NK⁺Caspase-8, the curative effect and adverse events.ResultAfter treatment, PLT amount in both groups increased, and the increase in the observation group was much higher than that of the control group, the difference had statistical significance (P < 0.05). The time needed for PLT amount in the control group to reach the normal and peak values was longer than that of the observation group, whose PLT peak value was higher than another group. The difference had statistical significance (P < 0.05). After the treatment, the levels of TNF-α and IL-6 were lowered, and the value of the observation group was lower than that of another. The difference between and within the group has statistical significance. After the treatment, the level of CD3⁺, CD3⁺CD95L⁺ and CD3⁺Caspase-8⁺ is much higher and CD3⁺Caspase-3⁺ level lower than that before the treatment. The difference has statistical significance (P < 0.05). After the treatment, the level of NK⁺ and NK⁺ CD95L⁺ is higher and the level of NK⁺Caspase-8⁺ lower than that before the treatment. The difference has statistical significance (P < 0.05). After the treatment, the total effective rate 91.67% of the observation group is much higher than that 75.56% of another. The difference has statistical significance (P < 0.05). After the treatment, the incidence rate of adverse events in the control group is 11.11% (5/45), while 4.17% (2/48) in the observation group. The difference between groups has statistical significance (χ² = 3.890, P < 0.05).ConclusionThe curative effect of Prednisone in combination with Mycophenolate on ITP patients is better than orally taking Prednisone tablets. Moreover, when it comes to Prednisone in combination with Mycophenolate, both the PLT amount and immunocompetence are improved without much adverse reaction, and the molecules of peripheral blood T lymphocytes and NK lymphocytes can be effectively adjusted to relieve the symptoms. So the method is trustworthy to be popularized for clinical practices.     

Co-administration of plasmid-encoded granulocyte-macrophage colony-stimulating factor increases human immunodeficiency virus-1 DNA vaccine-induced polyfunctional CD4+ T-cell responses

T-cell based vaccines against human immunodeficiency virus (HIV) generate specific responses that may limit both transmission and disease progression by controlling viral load. Broad, polyfunctional, and cytotoxic CD4⁺T-cell responses have been associated with control of simian immunodeficiency virus/HIV-1 replication, supporting the inclusion of CD4⁺ T-cell epitopes in vaccine formulations. Plasmid-encoded granulocyte-macrophage colony-stimulating factor (pGM-CSF) co-administration has been shown to induce potent CD4⁺ T-cell responses and to promote accelerated priming and increased migration of antigen-specific CD4⁺ T-cells. However, no study has shown whether co-immunisation with pGM-CSF enhances the number of vaccine-induced polyfunctional CD4⁺ T-cells. Our group has previously developed a DNA vaccine encoding conserved, multiple human leukocyte antigen (HLA)-DR binding HIV-1 subtype B peptides, which elicited broad, polyfunctional and long-lived CD4⁺ T-cell responses. Here, we show that pGM-CSF co-immunisation improved both magnitude and quality of vaccine-induced T-cell responses, particularly by increasing proliferating CD4⁺ T-cells that produce simultaneously interferon-γ, tumour necrosis factor-α and interleukin-2. Thus, we believe that the use of pGM-CSF may be helpful for vaccine strategies focused on the activation of anti-HIV CD4⁺ T-cell immunity.     

结直肠癌围术期营养干预对患者术后营养指标、T淋巴细胞亚群及并发症的影响

目的:探讨结直肠癌围术期营养干预对患者术后营养指标、T淋巴细胞亚群及并发症的影响。方法:选取2017年3月～2019年3月期间我院收治的结直肠癌患者103例,根据随机数字表法将患者分为对照组(n=51)和研究组(n=52),对照组给予常规方案干预,研究组给予营养支持方案干预。比较两组围术期指标、营养指标、T淋巴细胞亚群及并发症。结果:研究组患者的排便时间、首次肛门排气时间、住院时间均短于对照组(P0.05)。对照组术后1 d、术后7 d的CD3+、CD4~+、CD4~+/CD8~+均低于入院时,CD8~+则高于入院时(P0.05);研究组术后1 d、术后7 d的CD3~+、CD4~+、CD4~+/CD8~+呈先降低后升高趋势,CD8~+呈先升高后降低趋势(P0.05);研究组术后1 d、术后7 d的CD3+、CD4~+、CD4~+/CD8~+高于对照组,CD8~+则低于对照组(P0.05)。两组术后1 d、术后7 d血清前清蛋白(PA)、血红蛋白(Hb)、总蛋白(TP)、转铁蛋白(TF)水平均呈先下降后升高趋势(P0.05);研究组术后1 d、术后7 d血清PA、Hb、TP、TF水平均高于对照组(P0.05)。两组并发症发生率比较差异无统计学意义(P0.05)。结论:结直肠癌围术期给予营养支持方案干预,可有效改善患者营养状态,提高免疫功能,且不增加并发症发生率,具有一定的临床应用价值。     

Effect of a subset of adipose-derived stem cells isolated with liposome magnetic beads to promote cartilage repair

This study aimed to investigate the ability of CD146⁺ subset of ADSCs to repair cartilage defects. In this study, we prepared CD146⁺ liposome magnetic beads (CD146⁺LMB) to isolate CD146⁺ADSCs. The cells were induced for chondrogenic differentiation and verified by cartilage-specific mRNA and protein expression. Then a mouse model of cartilage defect was constructed and treated by filling the induced cartilage cells into the damaged joint, to evaluate the function of such cells in the cartilage microenvironment. Our results demonstrated that the CD146⁺LMBs we prepared were uniform, small and highly stable, and cell experiments showed that the CD146⁺LMB has low cytotoxicity to the ADSCs. ADSCs isolated with CD146⁺LMB were all CD146⁺, CD105⁺, CD166⁺ and CD73⁺. After chondrogenic induction, the cells showed significantly increased expression of cartilage markers Sox9, collagen Ⅱ and aggrecan at protein level and significantly increased Sox9, collagen Ⅱ and aggrecan at mRNA level, and the protein expression and mRNA expression of CD146⁺ADSCs group were higher than those of ADSCs group. The CD146⁺ADSCs group showed superior tissue repair ability than the ADSCs group and blank control group in the animal experiment, as judged by gross observation, histological observation and histological scoring. The above results proved that CD146⁺LMB can successfully isolate the CD146⁺ADSCs, and after chondrogenic induction, these cells successfully promoted repair of articular cartilage defects, which may be a new direction of tissue engineering.     

腹腔镜与开腹手术对进展期胃癌患者围手术期疗效及免疫功能的影响

目的:研究腹腔镜与开腹手术对进展期胃癌患者围手术期疗效及免疫功能的影响。方法:选择从2015年9月到2016年8月在医院治疗的进展期胃癌患者124例作为研究对象,依照简单数字随机表法将患者分成观察组以及对照组各含62例,对观察组进行腹腔镜辅助手术,对照组则给予开腹手术,对比两组围术期疗效相关指标,手术前后的免疫功能指标CD4~+、CD8~+及CD4~+/CD8~+),以及两组术后的并发症。结果:观察组的手术时间明显长于对照组,且切口长度、出血量少于对照组,住院时间和术后排气时间均较对照组短,差异有统计学意义(P0.05)。术后两组CD4~+、CD4~+/CD8~+水平均较术前明显降低,而CD8~+水平明显上升,且术后观察组CD4~+、CD4~+/CD8~+水平均显著高于对照组,差异有统计学意义(P0.05)。术后观察组并发症的总发生率是4.84%,明显低于对照组的16.13%,差异有统计学意义(P0.05)。结论:利用腹腔镜手术治疗进展期胃癌,具有较好的疗效,且对患者免疫功能产生的影响较小,安全性较高,值得给予推广。     

Selective depletion of FOXP3high cells by Fas–Fas-L–induced apoptosis occurs in CD4+CD25+-enriched populations during repeated expansion

Background aimsExpansion of anti-CD25 bead-isolated human Tregs culture has paradoxically resulted in reduced suppressive activity, but the mechanism(s) responsible for these observations are poorly defined.MethodsMagnetic-bead isolated human CD25⁺ cells were expanded with anti-CD3/CD28 beads and high doses of rhIL-2. Detection of Fas and Fas ligand (Fas-L) expression, activation of Caspase 8, cell proliferation and cytokine production was evaluated by multi-color fluorescence-activated cell sorting analysis. The role of Fas–Fas-L–mediated cell death was dissected through the use of agonist or antagonist monoclonal antibodies directed at Fas and Fas-L.ResultsRepeated expansion of bead-enriched CD4⁺CD25⁺ cells generated a cellular product with markedly reduced suppressive activity and with significantly increased CD8⁺ T cells and CD4⁺ T cells producing interferon-γ and/or interleukin-2. We showed that Fas–Fas-L–mediated apoptosis of CD4⁺FOXP3^high cells and rapid cell-cycling of CD8⁺ T cells were collectively responsible for the reduced proportion of CD4⁺FOXP3^high cells in expanded cultures. The depletion of CD4⁺FOXP3^high cells and activation of Caspase 8 in CD4⁺FOXP3^high cells was attenuated by Fas antagonist antibody, ZB4, in short-term culture. However, the loss of CD4⁺FOXP3^high cells during expansion was not prevented by either Fas or Fas-L antagonist antibodies.ConclusionsTaken together, the data show that Fas–Fas-L–mediated apoptosis may limit the expansion of anti-CD25 bead-isolated cells in vitro.