Induction of apoptosis in human colon carcinoma cells HT29 by sublethal cryo-injury: mediation by cytochrome c release

Cryosurgery is an emerging treatment for human solid tumors, notably colorectal liver metastasis. Cryosurgical procedures generate a thermal gradient of from at least -50 degrees C at the center of the tumor being treated to about 0 degrees C at the periphery. Cell death occurs by necrosis in the center, while the peripheral zone of frozen tumor harbors a mix of viable and dead tissue. In order to understand the mechanisms of cell death and survival in this peripheral area at risk for tumor recurrence, we have established an in vitro freezing system that mimics in vivo conditions of sublethal injury. HT29 colon cancer cells were subjected to freezing temperatures from -6 degrees C to -36 degrees C, thawed at room temperature for 30 min and rewarmed at 37 degrees C for a period of time. Post-freeze-thaw, cryolytic cells were evaluated by trypan blue exclusive assay. We also identified apoptotic cells after rewarming by cell shrinkage, nucleic condensation, TUNEL assay, DNA fragmentation and PARP degradation. The intensity of cryolysis and apoptosis was increased by lowering the freezing temperature. At -36 degrees C, all cells were dead immediately after freeze-thaw. A kinetic analysis of cryo-induced apoptosis showed that the commitment to enter apoptosis occurred right after the freeze-thaw period and lasted less than 8 hr after rewarming. We further demonstrated that freezing triggers one of the caspase cascade involved in apoptosis: release of cytochrome c from mitochondria to cytosol, followed by activation of caspase-9 and degradation of PARP. These results indicate the death of cancer cells under cryo-treatment at sublethal freezing temperature can be attributed 2 different modes, cryolysis as well as apoptosis. HT29 cells carrying p53 mutant have very quick response for induction of apoptosis by cryo-treatment and contain an intact pathway of caspase cascade. Further studies will address if mechanisms in cells with wild-type p53 will differ.     

Effects of Maxingloushi decoction on immune inflammation and programmed death markers in mice with chronic obstructive pulmonary disease

BACKGROUND: To investigate effects of Maxingloushi decoction on lung inflammation and programmed death markers(programmed death-1 [PD-1], programmed death-ligand 1 [PD-L1]) in the lung tissue, peripheral blood, and bronchoalveolar lavage fl uid(BLF) in a mouse model of chronic obstructive pulmonary disease(COPD).METHODS: Thirty-six mature male BALB/C mice were randomly divided into normal group(group A, n=6), COPD model group(group B, n=10), Maxingloushi decoction + COPD group(group C, n=10), and PD-1 inhibitor + COPD group(group D, n=10). The COPD model was established by smoke inhalation combined with lipopolysaccharide(LPS). Levels of PD-1 and PD-L1 in plasma and BLF were measured by enzyme-linked immunosorbent assay(ELISA). Histopathological techniques were used to semi-quantitatively analyze the immuno-fluorescence optical density(IOD) value of the lung tissue. RESULTS: In plasma and BLF, the expression of PD-1 in the group B was higher than that in the group A, and the expression of PD-L1 was lower than that in the group A. The expression of PD-1 and PD-L1 in the lung tissue was normalized in the group C in comparison with the group B(P<0.05) and the group D(P<0.05), and infl ammatory cell infiltration in the lung tissue was also improved.CONCLUSIONS: These findings reveal that COPD causes an immune imbalance in the peripheral blood and lung tissue, and that both Maxingloushi decoction and PD-1 inhibitor treatment can mitigate lung inflammation in COPD by reducing PD-1 expression and increasing PD-L1 expression. The treatment effect of Maxingloushi decoction may be superior to that of PD-1 inhibitor.     

Effect of Stabilizer on the Maximum Degree and Extent of Supersaturation and Oral Absorption of Tacrolimus Made By Ultra-Rapid Freezing

Purpose

Solid dispersions containing various stabilizers and tacrolimus (TAC) prepared by an Ultra-rapid Freezing (URF) process were investigated to determine the effect on their ability to form supersaturated solutions in aqueous media and on enhancing transport across biological membranes.

Materials and Methods

The stabilizers included poly(vinyl alcohol; PVA), poloxamer 407 (P407), and sodium dodecyl sulfate (SDS). In vivo absorption enhancement in rats was also investigated. Dissolution studies were conducted at supersaturated conditions in both acidic media for 24 h and at delayed release (enteric) conditions to simulate intestinal transit.

Results

The rank order of C/Ceq_max in the dissolution studies at acidic conditions was URF-P407?>?URF-SDS?>?Prograf® (PRO)?>?URF-PVA:P407. For C/Ceq_max under enteric conditions, the order was URF-SDS?>?PRO?>?URF-PVA:P407?>?URF-P407, and for the extent of supersaturation (AUC) in acidic and pH shift conditions it was URF-SDS>PRO>URF-PVA:P407>URF-P407. The pharmacokinetic data suggests URF-P407 had the greatest absorption having higher C _max with a 1.5-fold increase in AUC compared to PRO. All URF compositions had a shorter T _max compared to PRO.

Conclusions

The nanostructured powders containing various stabilizing polymers formed by the URF process offer enhanced supersaturation characteristics leading to increased oral absorption of TAC.     

外周血PD⁃1/PD⁃L1 在乳腺癌中的研究进展

2019年阿替利珠单抗获批应用于无法手术的程序性死亡受体‐配体1（PD‐L1）阳性三阴型乳腺癌,并以组织中 PD‐L1阳性表达作为应用此种抑制剂的指征。现有研究已证明程序性死亡受体‐1（PD‐1）与PD‐L1的表达能够提示肿瘤的大 小、分级、分期、预后,反映机体的免疫状态以及对免疫抑制剂治疗的敏感性和疗效。但临床上仍然需要一种可重复性较高的 手段替代组织活检对机体免疫状态进行实时监测以随时调整临床用药。因此,寻找其他形式的PD‐1/PD‐L1检测替代组织病 理学检查成为研究重点。外周血的检测不失为一种替代方式。本文将就目前乳腺癌患者外周血中血浆可溶性PD‐1（sPD‐1） 与血浆可溶性PD‐L1（sPD‐L1）、外泌体PD‐1 与外泌体PD‐L1、循环肿瘤细胞（CTC）PD‐1 与PD‐L1 及循环淋巴细胞PD‐1 与 PD‐L1等不同形式PD‐1与PD‐L1检测的研究现状、实际意义及其与病理组织中PD‐1/PD‐L1表达性质的关联做了简要综述,并 指出乳腺癌中该领域的研究方向及面临的挑战,为临床PD‐1/PD‐L1检测新手段提供新的方向和思路。     

Daratumumab With Cetrelimab,an Anti–PD-1 Monoclonal Antibody,in Relapsed/Refractory Multiple Myeloma

BackgroundDaratumumab is approved for relapsed or refractory multiple myeloma (RRMM) as monotherapy or in combination regimens. We evaluated daratumumab plus cetrelimab, a programmed death receptor-1 inhibitor, in RRMM.Patients and MethodsThis open-label, multiphase study enrolled adults with RRMM with ≥ 3 prior lines of therapy. Part 1 was a safety run-in phase examining dose-limiting toxicities of daratumumab (16 mg/kg intravenously weekly for cycles 1-2, biweekly for cycles 3-6, and monthly thereafter) plus cetrelimab (240 mg intravenously biweekly, all cycles). In Parts 2 and 3, patients were to be randomized to daratumumab with or without cetrelimab (same schedule as Part 1). Endpoints included safety, overall response rate, pharmacokinetics, and biomarker analyses.ResultsNine patients received daratumumab plus cetrelimab in the safety run-in, and 1 received daratumumab in Part 2 before administrative study termination following a data monitoring committee’s global recommendation to stop any trial including daratumumab combined with inhibitors of programmed death receptor-1 or its ligand (programmed death-ligand 1). The median follow-up times were 6.7 months (safety run-in) and 0.3 months (Part 2). No dose-limiting toxicities occurred. All 10 patients had ≥ 1 treatment-emergent adverse event; 7 patients had grade 3 to 4 treatment-emergent adverse events, and none led to treatment discontinuation or death. In the safety run-in, 7 (77.7%) patients had ≥ 1 infusion-related reaction (most grade 1-2), and 1 had a grade 2 immune-mediated reaction. Among safety run-in patients, the overall response rate was 44.4%.ConclusionsNo new safety concerns were identified for daratumumab plus cetrelimab in RRMM. The short study duration and small population limit complete analysis of this combination.     

开胸术后镇痛效果的观察与研究

目的比较开胸术后两种镇痛方法的镇痛效果.方法将行开胸手术病人76例,随机分为观察组和对照组各38例.对照组采取病人静脉自控止痛法（PCIA）止痛,观察组采取肋间神经冷冻术止痛.比较观察组和对照组镇痛效果、手术前后24h心率、呼吸频率及动脉血气变化.结果2组镇痛效果、手术前后24 h心率、呼吸频率、动脉血气变化等观测指标的差异具有统计学意义（P＜0.05）.结论肋间神经冷冻术具有安全稳定的优势,是一种可靠的预防开胸术后剧烈疼痛的方法.     

Functional specialization of intestinal dendritic cell subsets during Th2 helminth infection in mice

Dendritic cells (DCs) are essential in dictating the nature and effectiveness of immune responses. In the intestine DCs can be separated into discrete subsets, defined by expression of CD11b and CD103, each with different developmental requirements and distinct functional potential. Recent evidence has shown that different intestinal DC subsets are involved in the induction of T helper (Th)17 and regulatory T cell responses, but the cells that initiate Th2 immune responses are still incompletely understood. We show that in the Th2 response to an intestinal helminth in mice, only CD11b⁺ and not CD11b^? DCs accumulate in the local lymph node, upregulate PDL2 and express markers of alternative activation. An enteric Th1 response instead activated both CD11b⁺ and CD11b^? DCs without eliciting alternative activation in either population. Functionally, only CD11b⁺ DCs activated during helminth infection supported Th2 differentiation in naive CD4⁺ T cells. Together our data demonstrate that the ability to prime Th2 cells during intestinal helminth infection, is a selective and inducible characteristic of CD11b⁺ DCs.     

Expression of programmed cell death 1/programmed cell death ligand 1 in the tumor microenvironments of primary gastrointestinal diffuse large B cell lymphomas

Background

Gastrointestinal diffuse large B cell lymphoma (GI DLBCL) is the most common gastrointestinal lymphoma. However, there has not been a comprehensive investigation into the expression patterns of programmed cell death 1 (PD-1) and programmed cell death ligand 1(PD-L1) in GI DLBCL tissues.

“冬病夏治”穴位液氮冷冻治疗慢性呼吸系统疾病的疗效观察

目的:观察"冬病夏治"穴位液氮冷冻治疗慢性呼吸系统疾病的疗效。方法:选取400例患者,经穴位液氮冷冻治疗,观察疗效。当年冬天未发病或发作明显减少,症状明显减轻视为显效;发作减少、症状减轻或发作较先前容易得到控制视为有效;治疗后症状、体征明显无改善,视为无效。结果:治疗后发病症状、程度明显有好转,确有疗效,且连续治疗时间越长,疗效越好。结论:"冬病夏治"穴位液氮冷冻治疗慢性呼吸系统疾病疗效确切,其作用机理有待系统研究。