Characterization and targeting ability evaluation of cell-penetrating peptide LyP-1 modified alginate-based nanoparticles

Doxorubicin hydrochloride (DOX) shows a powerful treatment effect on breast cancer. However, for its indiscriminate distribution after systemic administration, the therapeutic response of DOX will reduce and even result in serious adverse reactions during the long-term administration. To achieve better treatment, in this study we established a non-condensing sodium alginate-based nanoparticle-encapsulated DOX (DOX/NP), the surface of which was modified with cell-penetrating peptide LyP-1 (namely LyP-1-DOX/NP) to attain active targeting towards breast cancer cells. The size of LyP-1-DOX/NP was 138.50 ± 4.65 nm, with a polydispersity index (PDI) of 0.22 ± 0.02, and the zeta potential was 18.60 ± 0.49 mV. The drug loading efficiency (DLE) for the preparation was 91.21 ± 2.01%, with an encapsulation efficiency (EE) of 12.37 ± 0.35%. The nanoparticles exhibited good stability in vitro and slower release trend compared with free DOX in PBS at pH7.4. In vitro cytopharmacodynamics showed that LyP-1-DOX/NP had an excellent anti-breast cancer effect against MDA-MB-231 cells by the MTT test. The uptake amount of LyP-1-DOX/NP by MDA-MB-231 cells was much higher than that of free DOX or unmodified DOX/NP at all time points. Further in vivo pharmacokinetics studies showed that the concentration of LyP-1-DOX/NP was higher than that of free DOX or DOX/NP both in plasma and in tumor, suggesting its favorable long circulation and enhancing targeting property. The present study provides a promising strategy for using the LyP-1 cell-penetrating peptide to modify nanoparticles for enhancing their targeting ability towards breast cancer.

Doxorubicin hydrochloride (DOX) shows a powerful treatment effect on breast cancer.     

Two cases of brand-specific albumin sensitivity in patients receiving regular therapeutic plasma exchange

Therapeutic plasma exchange (PLEX) involves the removal of detrimental substances, commonly pathogenic antibodies or toxins, from a patient’s blood by exchanging their plasma with a replacement fluid. While a variety of replacement fluids are available, human albumin (4–5 %) is the most commonly used, as it is widely available, easily stored, and generally well tolerated. Despite its excellent safety profile, adverse reactions to albumin are well documented, ranging in severity from mild allergic symptoms to severe anaphylaxis. This report describes two cases of patients receiving frequent PLEX who developed sensitivities to human albumin. These patients differed substantially in the manifestations of their symptoms, the duration of their treatment, and their medical indication for PLEX. In both cases, symptom onset occurred shortly after completion of plasma exchange procedures and lasted for several hours. Symptoms disappeared when the patients were switched to albumin from a different manufacturer, suggesting that the reaction was specific to that formulation of albumin and not to the albumin itself. These cases highlight the possibility of manufacturer-specific acquired albumin sensitivities and provide a simple framework for the initial approach to the management of such reactions.     

IL-6抑制剂Sirukumab治疗类风湿关节炎疗效和安全性的meta分析

目的评价白细胞介素-6(interleukin-6,IL-6)抑制剂Sirukumab对类风湿关节炎(rheumatoid arthritis,RA)患者的疗效和安全性。方法计算机检索PubMed、Embase、Web of Science、Clinical Trials Website、Cochrane Library、中国知网(CNKI)、万方、维普等数据库中关于Sirukumab治疗RA有效性和安全性的随机对照试验(randomizedcontrolledtrials,RCTs),采用RevMan 5.3软件对数据进行meta分析。结果共纳入6篇RCTs,3197例患者。Meta分析结果显示,Sirukumab2个剂量组(50,100 mg)ACR20、ACR50、ACR70的有效率均高于安慰剂组(P<0.0001)。Sirukumab 2个剂量组不良反应、严重不良反应、严重感染、注射部位反应、因不良反应终止试验等的发生率高于对照组(P<0.05)。与阳性药Adalimumab比较,Sirukumab 2个剂量组DAS28-ESR的有效率较高(P<0.05),ACR50有效率差异无统计学意义(P>0.05)。结论现有证据表明Sirukumab治疗RA安全有效。受纳入文献数量的限制,上述结论还需进一步开展高质量的临床试验来评价其有效性和安全性。     

Analysis of CD8+ Treg cells in patients with ovarian cancer: a possible mechanism for immune impairment

Regulatory T (Treg) cells may participate in mediating a suppressive microenvironment that blunts successful anti-tumor immunotherapy. Recent studies show that CD8⁺ Treg cells might impede effective immune responses to established tumors. However, there is limited research regarding CD8⁺ Treg cells in ovarian cancer (OC) patients. Here, we investigated CD8⁺ Treg cells in OC patients and their in vitro induction. The immunohistochemistry of tumor-infiltrating lymphocytes revealed a significant correlation between the intratumoral CD8⁺ T cells and the forkhead box p3 (Foxp3)⁺ cells in the intraepithelial and stromal areas of advanced OC tissues. We examined the expression of Treg markers in CD8⁺ T cells from the peripheral blood and fresh tumor tissues of OC patients using flow cytometry. Our results indicated an increase in the CD8⁺ Treg cell subsets of OC patients compared with those in patients with benign ovarian tumors and healthy controls, including an increased expression of CD25, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), and Foxp3 and decreased CD28 expression. To demonstrate whether the tumor microenvironment could convert CD8⁺ effector T cells into suppressor cells, we used an in vitro transwell culturing system. Compared with the CD8⁺ T cells cultured alone, the CD8⁺ Treg cells induced in vitro by coculture with SK-OV-3/A2780 showed increased CTLA-4 and Foxp3 expression and decreased CD28 expression. In addition, the in vitro-induced CD8⁺ Treg cells inhibited naïve CD4⁺ T-cell proliferation, which was partially mediated through TGF-β1 and IFN-γ. Our study suggests that CD8⁺ Treg cells were increased in OC patients and could be induced in vitro, which may be the way that tumors limit antitumor immunity and evade immune surveillance.     

Diagnostic efficacy of three suction techniques for endoscopic ultrasound-guided fine-needle biopsy of solid pancreatic lesions: protocol for a multicenter randomized cross-over clinical trial

BackgroundHow suction technique affects endoscopic ultrasound-guided tissue acquisition (EUS-TA) remains unclear. A standardized protocol is currently lacking, with most previous studies being restricted to EUS-guided fine-needle aspiration (EUS-FNA). The research related to EUS-guided fine-needle biopsy (EUS-FNB) is sparse.AimsThe aim of this study is to evaluate the diagnostic efficacy, cellularity, tissue acquisition, blood contamination and adverse event rate of three common suction techniques (standard suction, slow-pull, and wet suction) used for EUS-FNB of solid pancreatic masses.MethodsThis is a multicenter single-blind randomized cross-over superiority trial. A total of 300 patients with suspected pancreatic malignancy will be enrolled from digestive endoscopic centers at five large tertiary hospitals in China. All three suction techniques will be performed on each patient using a 25G ProCore needle, with the sequence of suction techniques determined by randomization. Cytological and histological specimens obtained with each of the three techniques will be assessed independently. Outcomes among the three suction techniques will be compared.DiscussionTo the best of our knowledge, this is the largest multicenter randomized cross-over trial designed to determine the optimal suction technique for the diagnosis of solid pancreatic masses. This study may contribute to standardizing the suction technique for EUS-FNB.     

Intrahepatic Glissonian approach and outflow vascular occlusion during partial hepatectomy

Under ultrasound guidance, a blunt suture needle was inserted around the Glissonian pedicle and then sutured. This technique significantly reduced the blood loss and facilitated the procedure of partial hepatectomy. We applied this technique in 182 patients who needed partial hepatectomy. We concluded that this method is simple and easy to occlude the vascular inflow and outflow, and allows an accurate delineation of the anatomic zone and therefore, simplifies the procedure of partial hepatectomy.     

欧前胡素对小鼠急性肺损伤的影响及其机制研究

目的 探讨欧前胡素对小鼠急性肺损伤的影响及其作用机制。方法 将32只雌性BALB/c小鼠随机分为正常对照组、模型组、欧前胡素30?mg/kg组、欧前胡素60?mg/kg组。用欧前胡素预处理,脂多糖诱导进行模型复制,7?h后收集小鼠肺组织。测量肺湿/干重比,采用苏木精-伊红（HE）染色观察小鼠肺组织病理学改变,检测小鼠肺组织中髓过氧化物酶（MPO）活力及活性氧类（ROS）的水平,酶联免疫吸附试验（ELISA）检测小鼠肺组织肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）,Western blotting检测小鼠肺组织中磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B（Akt）、核转录因子κB（NF-κB）p65及基质金属蛋白酶-9 （MMP-9）蛋白表达水平。结果 与正常对照组比较,模型组小鼠肺组织损伤评分、肺组织ROS水平及PI3K、Akt、NF-κB p65蛋白磷酸化水平升高（P?<0.05）,TNF-α、IL-1β的释放及MMP-9蛋白表达水平升高（P?<0.05）;与模型组比较,欧前胡素组能够减轻肺组织损伤,降低肺损伤评分（P?<0.05）,降低小鼠肺组织中ROS水平及PI3K、Akt、NF-κB p65蛋白磷酸化水平（P?<0.05）,降低TNF-α、IL-1β的释放及MMP-9蛋白表达水平（P?<0.05）;与欧前胡素30?mg/kg组比较,欧前胡素60?mg/kg组小鼠肺损伤评分、肺组织ROS水平、Akt及NF-κB p65蛋白磷酸化水平、TNF-α的释放及MMP-9蛋白表达水平降低（P?<0.05）,PI3K蛋白磷酸化水平及IL-1β的释放差异无统计学意义（P?>0.05）。结论 欧前胡素对脂多糖诱导的急性肺损伤有保护作用,其作用机制可能与脂多糖诱导的急性肺损伤中ROS被抑制,以及ROS介导的PI3K/Akt/NF-κB通路被抑制有关。     

无症状性脑梗死与睡眠障碍共病的研究进展

无症状性脑梗死（SBI）是指常无明确的卒中史、典型的临床症状及神经系统阳性体征,需通过颅脑影像学或尸检发现的脑梗死。随着研究的深入,SBI已被证实并非完全无症状,部分患者可能存在头晕、头痛、睡眠障碍、记忆力下降等非特异性神经功能损害表现。同时,大量临床报道显示,睡眠障碍是脑卒中后常见的并发症,并常会对患者的预后和生活质量产生不利影响。SBI作为脑梗死类型中的一种,与睡眠障碍也存在一定的联系,但由于SBI的临床特征,本身常被临床医生忽视,睡眠问题便更不受关注,国内外也缺乏相关研究报道。因此,本文对SBI与睡眠障碍共病的可能发生机制以及可能伴发的负面影响进行了综述,以便提高临床医生对其的关注度,做到早期诊治。     

CD64 Expression Is Increased in Patients with Severe Acute Pancreatitis: Clinical Significance

Background/Aims

Upregulated CD64 expression on neutrophils is the most useful marker for acute bacterial infections and systemic inflammation. However, it is unknown whether CD64 is involved in the pathogenesis of acute pancreatitis (AP). This study was designed to determine whether CD64 is implicated in severe acute pancreatitis (SAP), and thus, is a suitable marker for SAP.

Methods

SAP was induced in rats with an intraperitoneal injection of L-arginine. CD64 expression in the rat pancreas was determined by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry. Additionally, the CD64 mRNA expression in peripheral blood leukocytes from 21 patients with mild acute pancreatitis (MAP) and 10 patients with SAP was investigated at the time of admission and during remission by qRT-PCR.

Results

CD64 mRNA and protein expression in the pancreas was significantly higher in rats with SAP, compared to the controls. The CD64 expression was higher in the patients with SAP than in the patients with MAP. During remission, CD64 mRNA decreased in both the MAP and SAP patients. The area under the curve of CD64 expression for the detection of SAP was superior to both the Ranson and the Acute Physiology and Chronic Health Evaluation II scores.

Conclusions

The CD64 level was significantly increased in correlation with the disease severity in SAP and may act as a useful marker for predicting the development of SAP.     

和厚朴酚对哮喘小鼠肺组织炎症反应的干预作用及其机制

目的：探讨和厚朴酚（HNK）对哮喘小鼠肺组织炎症反应的影响,阐明其干预作用及相关机制。方法：选取健康雌性C57BL/6J小鼠20只,随机分为对照组、模型组、地塞米松（DXM）组和HNK组,每组5只。采用鸡卵清蛋白（OVA）联合氢氧化铝[Al（OH）₃]悬液腹腔注射致敏和OVA滴鼻激发构建哮喘小鼠模型,在激发前30 min分别对DXM组和HNK组小鼠予以腹腔注射DXM和HNK溶液,模型组小鼠予以等量生理盐水。处死小鼠后取小鼠肺组织,HE染色观察评估肺组织病理形态表现;收集支气管肺泡灌洗液（BALF）,行快速迪夫染色,观察炎性细胞浸润程度;ELISA法检测各组小鼠血清中白细胞介素4（IL-4）、白细胞介素6（IL-6）和白细胞介素17（IL-17）水平,比色法检测各组小鼠肺组织匀浆中丙二醛（MDA）水平和超氧化物歧化酶（SOD）及谷胱甘肽过氧化物酶（GSH-Px）活性,Western blotting法检测各组小鼠肺组织中磷酸化c-Jun氨基末端激酶（p-JNK）、核因子κB（NF-κB）、含半胱氨酸的天冬氨酸蛋白水解酶（Caspase-3）、B淋巴细胞瘤-2（Bcl-2）和H2AX组蛋白（γH2Ax）表达水平,检测各组小鼠肺组织中γH2Ax免疫荧光强度。结果：与对照组比较,模型组小鼠气道上皮细胞脱落坏死数量明显增多,气道管壁增厚及气道旁炎症细胞浸润程度明显增加;与模型组比较,DXM组和HNK组小鼠上述表现减轻,而HNK组和DXM组小鼠肺组织病理改变表现相似。与对照组比较,模型组小鼠血清中IL-4、IL-6和IL-17水平升高（P<0.05）;与模型组比较,DXM组和HNK组小鼠血清中IL-4、IL-6和IL-17水平降低（P<0.05）;而HNK组小鼠血清中IL-4、IL-6和IL-17水平与DXM组比较差异无统计学意义（P>0.05）。与对照组比较,模型组小鼠肺组织匀浆中MDA水平升高（P<0.05）,而SOD和GSH-Px活性降低（P<0.05）;与模型组比较,DXM组和HNK组小鼠肺组织匀浆中MDA水平降低（P<0.05）,而SOD和GSH-Px活性升高（P<0.05）;HNK组小鼠MDA水平和SOD及GSH-Px活性与DXM组比较差异无统计学意义（P>0.05）。与对照组比较,模型组小鼠肺组织中p-JNK、NF-κB、Caspase-3和γH2Ax蛋白表达水平升高（P<0.05）,而Bcl-2蛋白表达水平降低（P<0.05）;与模型组比较,DXM组和HNK组小鼠肺组织中p-JNK、NF-κB、Caspase-3和γH2Ax蛋白表达水平降低（P<0.05）,而Bcl-2蛋白表达水平升高（P<0.05）;HNK组小鼠肺组织中p-JNK、NF-κB、Caspase-3、γH2Ax和Bcl-2蛋白表达水平与DXM组比较差异均无统计学意义（P>0.05）。与对照组比较,模型组小鼠肺组织中γH2Ax免疫荧光强度明显增强;与模型组比较,DXM组和HNK组小鼠肺组织中γH2Ax免疫荧光强度减弱,HNK组小鼠γH2Ax免疫荧光强度与DXM组接近。结论：HNK可在一定程度上抑制哮喘小鼠的肺组织损伤及炎症反应,其机制可能与其抑制氧化应激反应及DNA损伤有关。