二陈汤加减联合微波消融术对晚期非小细胞肺癌患者Th17/Treg失衡和血管生成因子的影响

摘要 目的：探讨二陈汤加减联合微波消融术（MWA）对晚期非小细胞肺癌（NSCLC）患者辅助性T细胞17（Th17）/调节性T细胞（Treg）失衡和血管生成因子的影响。方法：选择2020年9月~2022年9月期间我院收治的120例不可或不愿手术切除或体部立体定向放疗的ⅢB-Ⅳ期并同意MWA治疗的NSCLC患者。采用随机对照的方法将所有患者分为观察组（二陈汤加减联合MWA治疗，60例）及对照组（MWA治疗，60例）。对比两组中医主要证候总评分、EORTC生命质量测定量表（EORTC QLQ-C30）评分、Th17/Treg失衡、血清肿瘤标志物、血管生成因子和不良反应发生率。结果：治疗后，观察组中医主要证候总评分、EORTC QLQ- C30评分低于对照组（P<0.05）。治疗后，观察组Th17细胞比例、Treg细胞比例低于对照组，Th17/Treg比值高于对照组（P<0.05）。治疗后，观察组癌胚抗原（CEA）、鳞状细胞癌相关抗原（SCC-Ag）和角化素蛋白片段19（Cyfra21-1）低于对照组（P<0.05）。治疗后，观察组血管内皮生长因子（VEGF）和碱性成纤维细胞生长因子（bFGF）低于对照组（P<0.05）。两组不良反应发生率组间对比未见差异（P>0.05）。结论：二陈汤加减联合MWA对晚期NSCLC患者，可有效降低血清肿瘤标志物水平，促进临床症状改善，调节Th17/Treg比例失衡和血管生成因子。     

The contribution of heme propionate groups to the conformational dynamics associated with CO photodissociation from horse heart myoglobin

Photoacoustic calorimetry and transient absorption spectroscopy were used to study conformational dynamics associated with CO photodissociation from horse heart myoglobin (Mb) reconstituted with either Fe protoporphyrin IX dimethylester (FePPDME), Fe octaethylporphyrin (FeOEP), or with native Fe protoporphyrin IX (FePPIX). The volume and enthalpy changes associated with the Fe-CO bond dissociation and formation of a transient deoxyMb intermediate for the reconstituted Mbs were found to be similar to those determined for native Mb (DeltaV1 = -2.5+/-0.6 ml mol(-1) and DeltaH1 = 8.1+/-3.0 kcal mol(-1)). The replacement of FePPIX by FeOEP significantly alters the conformational dynamics associated with CO release from protein. Ligand escape from FeOEP reconstituted Mb was determined to be roughly a factor of two faster (tau=330 ns) relative to native protein (tau=700 ns) and accompanying reaction volume and enthalpy changes were also found to be smaller (DeltaV2 = 5.4+/-2.5 ml mol(-1) and DeltaH2 = 0.7+/-2.2 kcal mol(-1)) than those for native Mb (DeltaV2 = 14.3+/-0.8 ml mol(-1) and DeltaH2 = 7.8+/-3.5 kcal mol(-1)). On the other hand, volume and enthalpy changes for CO release from FePPIX or FePPDME reconstituted Mb were nearly identical to those of the native protein. These results suggest that the hydrogen bonding network between heme propionate groups and nearby amino acid residues likely play an important role in regulating ligand diffusion through protein matrix. Disruption of this network leads to a partially open conformation of protein with less restricted ligand access to the heme binding pocket.     

Imaging and manipulating the structural machinery of living cells on the micro- and nanoscale

The structure, physiology, and fate of living cells are all highly sensitive to mechanical forces in the cellular microenvironment, including stresses and strains that originate from encounters with the extracellular matrix (ECM), blood and other flowing materials, and neighbouring cells. This relationship between context and physiology bears tremendous implications for the design of cellular micro-or nanotechnologies, since any attempt to control cell behavior in a device must provide the appropriate physical microenvironment for the desired cell behavior. Cells sense, process, and respond to biophysical cues in their environment through a set of integrated, multi-scale structural complexes that span length scales from single molecules to tens of microns, including small clusters of force-sensing molecules at the cell surface, micron-sized cell-ECM focal adhesion complexes, and the cytoskeleton that permeates and defines the entire cell. This review focuses on several key technologies that have recently been developed or adapted for the study of the dynamics of structural micro-and nanosystems in living cells and how these systems contribute to spatially-and temporally-controlled changes in cellular structure and mechanics. We begin by discussing subcellular laser ablation, which permits the precise incision of nanoscale structural elements in living cells in order to discern their mechanical properties and contributions to cell structure. We then discuss fluorescence recovery after photobleaching and fluorescent speckle microscopy, two live-cell fluorescence imaging methods that enable quantitative measurement of the binding and transport properties of specific proteins in the cell. Finally, we discuss methods to manipulate cellular structural networks by engineering the extracellular environment, including microfabrication of ECM distributions of defined geometry and microdevices designed to measure cellular traction forces at micron-scale resolution. Together, these methods form a powerful arsenal that is already adding significantly to our understanding of the nanoscale architecture and mechanics of living cells and may contribute to the rational design of new cellular micro-and nanotechnologies.     

Quantification of the effect of electrical and thermal parameters on radiofrequency ablation for concentric tumour model of different sizes

Radiofrequency ablation (RFA) has been increasingly used in treating cancer for multitude of situations in various tissue types. To perform the therapy safely and reliably, the effect of critical parameters needs to be known beforehand. Temperature plays an important role in the outcome of the therapy and any uncertainties in temperature assessment can be lethal. This study presents the RFA case of fixed tip temperature where we’ve analysed the effect of electrical conductivity, thermal conductivity and blood perfusion rate of the tumour and surrounding normal tissue on the radiofrequency ablation. Ablation volume was chosen as the characteristic to be optimised and temperature control was achieved via PID controller. The effect of all 6 parameters each having 3 levels was quantified with minimum number of experiments harnessing the fractional factorial characteristic of Taguchi’s orthogonal arrays. It was observed that as the blood perfusion increases the ablation volume decreases. Increasing electrical conductivity of the tumour results in increase of ablation volume whereas increase in normal tissue conductivity tends to decrease the ablation volume and vice versa. Likewise, increasing thermal conductivity of the tumour results in enhanced ablation volume whereas an increase in thermal conductivity of the surrounding normal tissue has a debilitating effect on the ablation volume and vice versa. With increase in the size of the tumour (i.e., 2–3 cm) the effect of each parameter is not linear. The parameter effect varies with change in size of the tumour that is manifested by the different gradient observed in ablation volume. Most important is the relative insensitivity of ablation volume to blood perfusion rate for smaller tumour size (2 cm) that is also in accordance with the previous results presented in literature. These findings will provide initial insight for safe, reliable and improved treatment planning perceptively.     

Elevated serum levels of heat shock protein 70 can be detected after radiofrequency ablation

Due to their adjuvant effect and their ability to chaperone tumor-associated peptides, heat shock proteins constitute a potent alarm signal for the immune system and can lead to activation of anti-tumor T-cell immunity. Radiofrequency ablation has been reported to induce heat shock protein expression especially that of heat shock protein 70 in sublethally damaged tumor cells. In this study, we evaluated the release of heat shock protein 70 into the serum of cancer-bearing patients directly after radiofrequency ablation. Sera of 22 patients undergoing radiofrequency ablation for the treatment of primary and secondary malignancies of the liver, kidney, and lung, as well as control sera of 20 patients undergoing diagnostic liver biopsy were analyzed using a manufactured heat shock protein 70 ELISA. A significant increase in serum levels of heat shock protein 70 was detectable in the patient cohort 1 day after radiofrequency ablation. More than a twofold increase was observed in nine out of 22 patients, which tended to correlate with favorable clinical outcome. No patient of the control group revealed a comparable increase. Radiofrequency ablation can lead to a release of heat shock protein 70 into the serum, which is transiently detectable 1 day after treatment. Elevated heat shock protein 70 serum levels may constitute a biomarker for favorable clinical outcome.     

Laser Ablation of the Zebrafish Pronephros to Study Renal Epithelial Regeneration

Acute kidney injury (AKI) is characterized by high mortality rates from deterioration of renal function over a period of hours or days that culminates in renal failure¹. AKI can be caused by a number of factors including ischemia, drug-based toxicity, or obstructive injury¹. This results in an inability to maintain fluid and electrolyte homeostasis. While AKI has been observed for decades, effective clinical therapies have yet to be developed. Intriguingly, some patients with AKI recover renal functions over time, a mysterious phenomenon that has been only rudimentally characterized^1,2. Research using mammalian models of AKI has shown that ischemic or nephrotoxin-injured kidneys experience epithelial cell death in nephron tubules^1,2, the functional units of the kidney that are made up of a series of specialized regions (segments) of epithelial cell types³. Within nephrons, epithelial cell death is highest in proximal tubule cells. There is evidence that suggests cell destruction is followed by dedifferentiation, proliferation, and migration of surrounding epithelial cells, which can regenerate the nephron entirely^1,2. However, there are many unanswered questions about the mechanisms of renal epithelial regeneration, ranging from the signals that modulate these events to reasons for the wide variation of abilities among humans to regenerate injured kidneys.The larval zebrafish provides an excellent model to study kidney epithelial regeneration as its pronephric kidney is comprised of nephrons that are conserved with higher vertebrates including mammals^4,5. The nephrons of zebrafish larvae can be visualized with fluorescence techniques because of the relative transparency of the young zebrafish⁶. This provides a unique opportunity to image cell and molecular changes in real-time, in contrast to mammalian models where nephrons are inaccessible because the kidneys are structurally complex systems internalized within the animal. Recent studies have employed the aminoglycoside gentamicin as a toxic causative agent for study of AKI and subsequent renal failure: gentamicin and other antibiotics have been shown to cause AKI in humans, and researchers have formulated methods to use this agent to trigger kidney damage in zebrafish^7,8. However, the effects of aminoglycoside toxicity in zebrafish larvae are catastrophic and lethal, which presents a difficulty when studying epithelial regeneration and function over time. Our method presents the use of targeted cell ablation as a novel tool for the study of epithelial injury in zebrafish. Laser ablation gives researchers the ability to induce cell death in a limited population of cells. Varying areas of cells can be targeted based on morphological location, function, or even expression of a particular cellular phenotype. Thus, laser ablation will increase the specificity of what researchers can study, and can be a powerful new approach to shed light on the mechanisms of renal epithelial regeneration. This protocol can be broadly applied to target cell populations in other organs in the zebrafish embryo to study injury and regeneration in any number of contexts of interest.     

B 超引导下子宫内膜消融术治疗更年期功血51 例临床分析

目的:探讨在B超引导下子宫内膜消融术治疗更年期功血的效果。方法:扩张宫颈至6.5号,吸宫,在B超监视下由子宫右前壁始向外刮凝,依次顺时针方向刮凝宫腔2周。结果:治愈31例(60.8%),有效19例(37.3%),好转1例(1.9%),无效0例。结论:电凝刀子宫内膜消融术与其他更年期功血治疗方法相比,消融的范围及深度由B超监视,安全性高,创伤小,不开刀,恢复快,并且治愈率高,更容易在临床上推广。     

Production of CETD transgenic mouse line allowing ablation of any type of specific cell population

Diphtheria toxin A-chain (DT-A) is a potent inhibitor of protein synthesis. As little as a single molecule of DT-A can result in cell death. DT-A gene driven by a tissue-specific promoter is used to achieve genetic ablation of a particular cell lineage. However, this transgenic approach often results in aberrant depletion of unrelated cells. To avoid this, we established a method for specific depletion of a cell population by controlled expression of the DT-A gene via the Cre-loxP system. We produced five transgenic mice carrying CETD construct containing loxP-flanked enhanced green fluorescent protein (EGFP) cDNA and the DT-A gene. Transfection of primary cultured cells derived from CETD transgenic fetus with Cre expression plasmid resulted in extensive cell loss, as expected. Bigenic (double transgenic) offspring obtained by crossbreeding between CETD and MNCE transgenic mice in which Cre expression is controlled by the myelin basic protein (MBP) promoter exhibited embryonic lethality, suggesting expression of Cre at embryonic stages. Intravenous injection of Cre expression vector to CETD mice led to generation of glomerular lesions, probably due to predominant depletion of glomerular epithelial cells. This Cre-loxP-based cell ablation technology is powerful and convenient method of generating mice lacking any chosen cell population.     

人眼波前像差表示、测量及矫正研究

目的:波前像差引导的准分子激光角膜消融是屈光手术的新方法,研究人眼波前像差的测量原理、方法、表示、人眼波前像差准分子激光矫正的原理,以此理论用于准分子激光人眼像差矫正系统。方法:采用理论研究、计算机模拟、实验室实验等手段。分析人眼像差的概念和产生的原因,用数学的Zern ike多项式来表示像差,理论上定量分析Zern ike多项式表示的波前像差与角膜切削深度的关系,研究准分子激光切削角膜的机理,研究准分子激光进行矫正人眼像差的原理框图。结果:通过计算机模拟和实验室实验,用准分子激光矫正低阶和高阶像差是可行的。结论:用波前像差来引导屈光手术,使人眼的视力能够达到20/10上,并能避免当前PRK、LASIK屈光手术前后像差增大而引起的对视觉质量的影响。