Immunoprotective activity of a Salmonid Alphavirus Vaccine: comparison of the immune responses induced by inactivated whole virus antigen formulations based on CpG class B oligonucleotides and poly I:C alone or combined with an oil adjuvant

CpG oligonucleotides and polyinosinic:polycytidylic acid (poly I:C) are toll-like receptor (TLR) agonists that mimic the immunostimulatory properties of bacterial DNA and double-stranded viral RNA respectively, and which have exhibited potential to serve as vaccine adjuvants in previous experiments. Here, a combination of CpGs and poly I:C together with water- or oil-formulated Salmonid Alphavirus (SAV) antigen preparations has been used for a vaccine in Atlantic salmon and tested for protection in SAV challenge trial. The results demonstrate that vaccination with a high dose of the SAV antigen induced protection against challenge with SAV which correlated with production of neutralizing antibodies (NAbs). As the high antigen dose alone induced full protection, no beneficial effect from the addition of CpG and poly I:C could be observed. Nevertheless, these TLR ligands significantly enhanced the levels of NAbs in serum of vaccinated fish. Interestingly, gene expression analysis demonstrated that while addition of oil suppressed the CpG/poly I:C-induced expression of IFN-γ, the upregulation of IFNa1 was substantially enhanced. A low dose of the SAV antigen combined with oil did not induce any detectable levels of NAbs either with or without TLR ligands present, however the addition of CpG and poly I:C to the low SAV antigen dose formulation significantly enhanced the protection against SAV suggesting that CpG/poly I:C may have enhanced a cytotoxic response - a process which is dependent on the up-regulation of type I IFN. These results highlight the immunostimulatory properties of the tested TLR ligands and will serve as a ground for further, more detailed studies aimed to investigate their capacity to serve as adjuvants in vaccine formulations for Atlantic salmon.     

三种创面敷料在薄中厚皮片供皮区的应用研究

目的观察3种创面敷料对薄中厚皮片供皮区创面愈合的影响。 方法选取蚌埠医学院第一附属医院整形烧伤科2020年1月至12月收治的38例自体皮片移植术患者。在同一患者供皮区分别取相同面积的类矩形薄中厚皮片3处,每处均间隔1 cm,每例所取皮片总面积基本相同,将同一患者的3处供皮区分为凡士林敷料组、银离子藻酸盐敷料组和丝素蛋白膜状敷料组3组,取皮后分别贴敷凡士林敷料、银离子藻酸盐敷料和丝素蛋白膜状敷料。对比3组供皮区创面积血率、初次换药时患者的疼痛程度[数字评定量表(NRS)]、创面感染率、创面上皮化愈合时间、创面后期愈合效果。对数据行单因素方差分析、t检验和χ²检验。 结果(1)创面积血率：丝素蛋白膜状敷料组创面积血率(23.68%)分别高于分别高于凡士林敷料组(2.63%)和银离子藻酸盐敷料组(5.26%),差异均有统计学意义(χ²＝ 7.370、5.208, P<0.05),凡士林敷料组与银离子藻酸盐敷料组积血率比较,差异无统计学意义(χ²＝0.347, P>0.05);(2)初次换药时疼痛程度评价：丝素蛋白膜状敷料组的NRS评分为(2.97±1.48)分,分别低于银离子藻酸盐敷料组[(3.97±1.84)分]和凡士林敷料组[(6.03±1.37)分],差异均有统计学意义(t＝ 4.854、0.873, P<0.05);银离子藻酸盐敷料组疼痛评分低于凡士林敷料组,差异有统计学意义(t＝1.467, P<0.05);(3)创面感染率：银离子藻酸盐敷料组创面感染率(5.26%)分别与丝素蛋白膜状敷料组(0)和凡士林敷料组(10.53%)比较,差异均无统计学意义(χ²＝ 2.054、0.724, P>0.05);丝素蛋白膜状敷料组与凡士林敷料组比较,感染率低,差异有统计学意义(χ²＝ 4.222, P<0.05);(4)创面上皮化愈合时间：丝素蛋白膜状敷料组创面上皮化愈合时间为(8.95±1.34) d,与银离子藻酸盐敷料组[(13.69±1.64) d]以及凡士林敷料组[(11.78±1.43) d]比较,愈合时间均较短,差异均有统计学意义(t＝0.953、1.204, P<0.05)。与银离子藻酸盐敷料组比较,凡士林敷料组愈合时间短,差异有统计学意义(t＝2.147, P<0.05);(5)创面后期愈合效果：3组在瘢痕增生和色素沉着2方面均无明显差异。 结论丝素蛋白膜状敷料应用于薄中厚皮片供皮区,具有相对无痛、抗感染能力强、上皮化愈合时间短等优势,但在防止创面积血方面效果欠佳。     

Ag Nanocluster-Enhanced Scintillation Properties of Borophosphate Glasses Doped with CsPbBr3 Quantum Dots

A novel and effective method to improve scintillation properties of glass-ceramics, such as intensity enhancement and decay-time shortening, is reported in this work. Compared with crystal scintillators, glass scintillators always have the problems of low efficiency and long decay; how to solve them has always been a scientific puzzle in the field of scintillation glass-ceramics. The plasma enhancement effect can be predicted to solve the above problems. Ag⁺ ions were diffused into glasses by ion exchange, and then Ag nanoparticles and CsPbBr₃ quantum dots were formed by heat treatment. The structure of the CsPbBr₃ perovskite consists of a series of shared corner PbBr₆ octahedra with Cs ions occupying the cuboctahedral cavities. By using Ag and the plasma resonance effect, the photoluminescence intensity of CsPbBr₃ quantum dot glasses was enhanced by 3 times, its radioluminescence intensity increased by 6.25 times, and its decay time was reduced by a factor of more than one. Moreover, the mechanism of photoluminescence and radioluminescence enhanced by Ag and plasma was discussed based on the experimental results and finite-difference time-domain method. We concluded that the increase in radioluminescence intensity was related to plasma enhancements and the energy exchange between Ag nanoclusters and CsPbBr₃ quantum dots. Doping Ag is a valid means to improve the scintillation luminescence of CsPbBr₃ quantum dot glasses, which can be applied in the field of scintillation.     

Nanofunctionalization of Additively Manufactured Titanium Substrates for Surface-Enhanced Raman Spectroscopy Measurements

Powder bed fusion using a laser beam (PBF-LB) is a commonly used additive manufacturing (3D printing) process for the fabrication of various parts from pure metals and their alloys. This work shows for the first time the possibility of using PBF-LB technology for the production of 3D titanium substrates (Ti 3D) for surface-enhanced Raman scattering (SERS) measurements. Thanks to the specific development of the 3D titanium surface and its nanoscale modification by the formation of TiO₂ nanotubes with a diameter of ~80 nm by the anodic oxidation process, very efficient SERS substrates were obtained after deposition of silver nanoparticles (0.02 mg/cm², magnetron sputtering). The average SERS enhancement factor equal to 1.26 × 10⁶ was determined for pyridine (0.05 M + 0.1 M KCl), as a model adsorbate. The estimated enhancement factor is comparable with the data in the literature, and the substrate produced in this way is characterized by the high stability and repeatability of SERS measurements. The combination of the use of a printed metal substrate with nanofunctionalization opens a new path in the design of SERS substrates for applications in analytical chemistry. Methods such as SEM scanning microscopy, photoelectron spectroscopy (XPS) and X-ray diffraction analysis (XRD) were used to determine the morphology, structure and chemical composition of the fabricated materials.     

Detection of Chlorpyrifos Using Bio-Inspired Silver Nanograss

Chlorpyrifos (CPF) is widely used as an organophosphorus insecticide; however, owing to developmental neurotoxicity, genotoxicity, and other adverse effects, it is harmful not only to livestock but also to humans. Therefore, the use of CPF was recently regulated, and its sensitive detection is crucial, as it causes serious toxicity, even in the case of residual pesticides. Because it is hard to detect the chlorpyrifos directly using spectroscopy (especially in SERS) without chemical reagents, we aimed to develop a SERS platform that could detect the chlorpyrifos directly in the water. In this study, we utilized the intrinsic properties of natural lawns that grow randomly and intertwine with each other to have a large surface area to promote photosynthesis. To detect CPF sensitively, we facilitated the rapid fabrication of biomimetic Ag nanograss (Ag-NG) as a surface-enhanced Raman spectroscopy (SERS) substrate using the electrochemical over-deposition method. The efficiency of the SERS method was confirmed through experiments and finite element method (FEM)-based electromagnetic simulations. In addition, the sensitive detection of CPF was enhanced by pretreatment optimization of the application of the SERS technique (limit of detection: 500 nM). The Ag-NG has potential as a SERS platform that could precisely detect organic compounds, as well as various toxic substances.     

Adjuvanted inactivated influenza A(H3N2) vaccines induce stronger immunogenicity in mice and confer higher protection in ferrets than unadjuvanted inactivated vaccines

Influenza viruses are major respiratory pathogens and the development of improved vaccines to prevent these infections is of high priority. Here, we evaluated split inactivated A(H3N2) vaccines (A/Uruguay/716/2007) combined or not with adjuvants (AS03, AS25 and Protollin) and administered by three different routes, intramuscular (i.m.), intranasal (i.n.) or intradermal (i.d.), both in BALB/c mice and in ferrets. Ferrets were challenged with the homologous strain A/Uruguay/716/2007 (H3N2) or the heterologous strain A/Perth/16/2009 (H3N2) 4 weeks after the second immunization with A/Uruguay/716/2007 vaccines. Temperature, weight loss and clinical signs were monitored on a daily basis and nasal washes were performed to evaluate viral titers in the upper respiratory tract. All adjuvanted vaccines induced stronger humoral immune responses than unadjuvanted ones in both mice and ferrets. In mice, the AS03- and AS25-adjuvanted i.m. vaccines generated a mixed Th1–Th2 response at 6 and 19 weeks after the last immunization as shown by the production of IgG1 and IgG2a antibodies as well as the production of IL-2, IL-4 and IFN-γ by CD4+ and CD8+ T cells. HAI and MN titers were also higher in those groups when compared to the i.n. Protollin-adjuvanted and unadjuvanted groups. The Protollin-adjuvanted i.n. vaccine induced a more Th1 oriented response with a significant production of IgA in bronchoalveolar lavages. In ferrets, the AS03- and AS25-adjuvanted i.m. vaccines also induced higher HAI and MN titers compared to the other groups. These vaccines also significantly decreased viral titers after challenge with both the homologous A/Uruguay/716/2007 (H3N2) and the heterologous A/Perth/16/2009 (H3N2) strains. In conclusion, adjuvanted influenza vaccines elicited stronger humoral response in mice and conferred greater protection in naive ferrets than unadjuvanted ones. Interestingly, the AS25 adjuvant system containing monophosphoryl-lipid-A appears particularly promising for developing more potent inactivated influenza vaccines.     

导向化疗对肿瘤细胞增殖动力学的影响

目的 :探讨导向化疗对肠癌细胞增殖动力学的影响。方法 :5 0例大肠腺癌患者随机分成三组 :A组单纯手术切除 ;B组术前静注化疗药物 ;C组术前静注导向化疗药物。术中取肿瘤组织及周围正常肠道组织进行AgNORs染色及检测。 结果 :1 手术组 (2 0例 )、化疗组 (13例 )、导向化疗组 (17例 )癌细胞AgNORs/核颗粒分别为4 18± 0 5 0、3 13± 0 2 0、2 2 9± 0 6 9,两两比较其差异存在显著性 (P <0 0 0 1) ;正常肠道细胞AgNORs/核均数分别为 1 81± 0 32、1 86± 0 2 5、1 80± 0 33,(P >0 0 5 )。 2 手术组、化疗组、导向化疗组肿瘤细胞AgNORs颗粒面积与细胞核面积的比值分别为 :0 2 85 8± 0 0 6 74、0 2 0 2 6± 0 0 5 93、0 2 0 0 9± 0 0 35 5 ,化疗组、导向化疗组与手术组比较其差异有显著性 (P <0 0 0 1) ,化疗组与导向化疗组之间的差异无显著性 (P >0 0 5 )。 3 手术组、化疗组、导向化疗组肿瘤细胞AgNORs颗粒光密度与肿瘤细胞核总光密度比值分别为 0 36 6 8± 0 0 6 0 3、0 30 36± 0 0 5 18、0 2 5 12± 0 0 374,两两比较差异有显著性 (P <0 0 1)。结论 :导向化疗与化疗均影响大肠癌细胞的增殖动力学 ,导向化疗对癌细胞增殖动力学的影响比化疗更明显。     

The Clinical Significance of Septin 9 and Colon Cancer Specific Antigen-2 (CCSA-2) in Colorectal Cancer

Introduction: Colorectal cancer (CRC) is a major health problem Worldwide, Egypt shows a high rate of early CRC in the world as 35% of 1,600 Egyptian CRC patients were under 40 with threefold increased risk of death within 5 years. DNA methylation-based biomarkers as methylated Septin9 (mSEPT9) has a promising role for detecting CRC. As well as set of nuclear matrix proteins associated with changes in the nuclear structure/architecture. detection of these nuclear proteins resulted in identification of biomarkers that are specific for colon cancer. Particular interest has been placed on colon cancer specific antigen-2(CCSA-2). Methods: A total of 30 newly diagnosed CRC patients, 7 colonic adenoma patients, and 15 age- and sex-matched control subjects were recruited in this study. Plasma mSEPT9was assayed by Epi procolon kit, CCSA-2 by ELISA and, Occult blood in stool by Guaiac-based fecal occult blood test. The level of Colon Cancer mSEPT9 and CCSA-2 were carried on CRC patients both preoperatively and three months postoperatively. Results: mSEPT9 has 96.7% sensitivity and 95.5% specificity in differentiating colorectal cancer patients from non-malignant cases. Also, our study showed a highly statistically significant difference between the pre and three months postoperative expression of mSEPT9 in colorectal cancer as there was a dramatically decrease in the expression of mSEPT9 postoperatively (p value < 0.001). The CCSA-2 at the cutoff level of >1.43 would provide 93.3% sensitivity and 90.9% specificity in differentiation between malignant and non-malignant cases. Also, the study showed that there is a statistically significant difference between colorectal cancer patients preoperatively and postoperatively according to CCSA-2 with dramatic decrease in its level postoperatively (p value > 0.001). Conclusion: The plasma SEPT9 DNA methylation level and Serum CCSA-2 could be used as promising non-invasive methods for observing the CRC patients postsurgical response to predict the occurrence of complete remission or relapses.     

当归注射液对溃疡性结肠炎患者血小板功能状态的影响及其意义

目的　探讨当归注射液对溃疡性结肠炎 (UC)患者血小板功能状态的影响及其治疗意义。方法　采用ELISA法 ,单克隆酶联免疫吸附法及血小板粘附检测仪检测 3 9例活动期及 2 5例缓解期溃疡性结肠炎患者与 3 0例健康对照者的血小板a颗粒膜蛋白 (GMP -14 0 )、血栓烷B2 (TXB2 )、6-酮 -前列腺素F1a( 6-Keto -PGF1a)、血小板粘附率及血管性假血友病因子相关抗原 (vWF :Ag)水平。结果　患者组血小板活化指标及血管内皮损伤程度显著高于对照组 (P <0 0 1) ,活动期UC患者又明显高于缓解期UC患者 (P <0 0 5 )。当归治疗组治疗前后血小板活化指标、血管内皮损伤指标及临床疗效有明显差异 (P <0 0 5 ) ,而一般治疗组治疗前后无明显差异 (P >0 0 5 )。结论　当归注射液能明显抑制UC患者体内血小板活化 ,减轻炎症反应 ,对溃疡性结肠炎有一定治疗价值。     

乙肝血清标志物-前S1抗原的临床意义

目的探讨乙肝病毒前-S1抗原与乙肝血清标记HBeAg、HBV-DNA的相互关系,了解乙肝病毒感染者血清前S1抗原的临床意义。方法双抗体夹心ELISA法测定血清前S1抗原,ELISA法测定乙肝病毒血清标记,荧光定量PCR检测血清乙肝病毒DNA。结果分析常规HBV血清标记物不同组合模式发现,HBeAg阳性模式中前S1抗原的检出率显著高于HBeAb阳性模式,其中S1抗原和HBeAg的符合率达80.88%;HBV-DNA阳性组前S1抗原的检出率明显高于HBV-DNA阴性组,两者的总体符合率达80.27%。结论血清前S1抗原与HBV-DNA、HBeAg相互关联,可以作为HBV存在和复制的标志。联合检测前S1抗原与HBV-DNA、HBeAg可相互补充、印证,对判断乙肝病毒的复制和存在更有价值。