A fluorescence method for homogeneous detection of influenza A DNA sequence based on guanine-quadruplex-N-methylmesoporphyrin IX complex and assistance-DNA inhibition

In his study, we report a fluorescence method for homogeneous detection of influenza A (H1N1) DNA sequence based on G-quadruplex-NMM complex and assistance-DNA (A-DNA) inhibition. The quadruplex-based functional DNA (QBF-DNA), composed of a complementary probe to the target H1N1 DNA sequence and G-rich fragment, was designed as the signal DNA. The A-DNA consisted of two parts, one part was complementary to target H1N1 DNA and the other part was complementary to the signal DNA. In the absence of target H1N1 DNA, the G-rich fragment of QBF-DNA can form G-quadruplex-NMM complex, which outputted a fluorescent signal. With the presence of target H1N1 DNA, QBF-DNA, and A-DNA can simultaneously hybridize with target H1N1 DNA to form double-helix structure. In this case, the A-DNA partially hybridized with the QBF-DNA, which inhibited the formation of G-quadruplex-NMM complex, leading to the decrease of fluorescent signal. Under the optimum conditions, the fluorescence intensity was inversely proportional to the concentration of target H1N1 DNA over the range from 25 to 700 pmol/L with a detection limit of 8 pmol/L. In addition, the method is target specific and practicability, and would become a new diagnostic assay for H1N1 DNA sequence and other infectious diseases.     

Prevalence and genetic diversity of human sapovirus associated with sporadic acute gastroenteritis in South China from 2013 to 2017

Human sapovirus (SaV) is an important viral agent for acute diarrhea worldwide, but timely prevalence data of human SaV in South China are still lacking. In this study, a 4-year surveillance was conducted to characterize the prevalence and genetic characteristics of the circulating SaV associated with sporadic diarrhea in South China. From November 2013 to October 2017, 569 fecal samples from patients with acute diarrhea were collected. SaV was detected in 11 samples with a positive rate of 1.93%. Three human genogroups of GI, GII, and GIV were identified, including five GI.1 strains, three GI.2 strains, one GI.3 strain, one GII.8 strain, and one GIV strain. Furthermore, multiple alignments of complete capsid protein VP1 genes of five local GI.1 strains and other available GI.1 strains in GenBank were performed. Average pairwise identities were calculated at 95.33% and 99.36% at nucleotide and amino acid levels, and only six variable amino acid sites were found during its 36-years’ evolution process. GI.1 strains could be further phylogenetically divided into four clusters with an approximate temporal evolution pattern, and local strains belonged to Cluster-d with other four strains from China and Japan. In summary, SaV was identified as an etiological agent responsible for sporadic gastroenteritis in Guangzhou with a low prevalence rate as in other Chinese cities, but its high genetic diversity suggested the necessity of continuous SaV surveillance in the future.     

Protective and therapeutic application of the depolymerase derived from a novel KN1 genotype of Klebsiella pneumoniae bacteriophage in mice

Klebsiella pneumoniae is one of the major Gram-negative bacterial pathogens causing hospital-acquired multidrug-resistant infections, and the antimicrobial treatment options are scarce. The lack of available antimicrobials has prompted the development of alternative strategies for the treatment of these infections. In this study, a K. pneumoniae bacteriophage (vB_KpnP_IME321) targeting a KN1 capsular type strain, Kp409, was isolated, characterized and sequenced. This bacteriophage has a latent period of 20 min and a burst size of approximately 410 pfu/cell. It contained 49 predicted open reading frames, of which ORF42 was identified as encoding the putative capsule depolymerase. The enzyme expressed and purified in the Escherichia coli BL21 system, namely Dp42, could depolymerize the capsular polysaccharide of Kp409 and form translucent halos on the plates. The phage-encoded depolymerase could increase the inhibitory effect of serum on the growth of bacteria in vitro. Pre-treated with Dp42 rescued 100% of mice following lethal Kp409 challenge, and administration of this enzyme after infection significantly increased survival rates of infected mice in the animal experiment. In conclusion, the phage-encoded depolymerase Dp42 represents a potential alternative strategy for controlling infections mediated by K. pneumoniae expressing the KN1 capsular polysaccharide.     

An Environmental Scan of the National and Provincial Diagnostic Reference Levels in Canada for Common Adult Computed Tomography Scans

Several regulatory bodies have agreed that low-dose radiation used in medical imaging is a weak carcinogen that follows a linear, non-threshold model of cancer risk. While avoiding radiation is the best course of action to mitigate risk, computed tomography (CT) scans are often critical for diagnosis. In addition to the as low as reasonably achievable principle, a more concrete method of dose reduction for common CT imaging exams is the use of a diagnostic reference level (DRL). This paper examines Canada's national DRL values from the recent CT survey and compares it to published provincial DRLs as well as the DRLs in the United Kingdom and the United States of America for the 3 most common CT exams: head, chest, and abdomen/pelvis. Canada compares well on the international scale, but it should consider using more electronic dose monitoring solutions to create a culture of dose optimization.     

载淫羊藿苷/凹凸棒石/Ⅰ型胶原/聚己内酯复合支架修复兔胫骨缺损的实验研究

目的探讨载淫羊藿苷/凹凸棒石/Ⅰ型胶原/聚己内酯（icariin/ attapulgite/collagen type Ⅰ/polycaprolactone，ICA/ATP/ColⅠ/PCL）复合支架修复兔胫骨缺损的效果。方法利用溶液铸浇-粒子滤沥法分别构建 ICA/20%ATP/ColⅠ/PCL（支架 1）、ICA/30%ATP/ColⅠ/PCL（支架 2）、20%ATP/ColⅠ/PCL（支架 3）、30%ATP/ColⅠ/PCL（支架 4）复合支架材料，采用扫描电镜观察支架 2 交联前、后结构特征，测量支架 2、4 的表面接触角检测材料吸水性能，采用体外降解实验评价支架 2 的药物缓释效果。取 30 只雄性日本大耳白兔，体质量（2.0±0.1）kg，随机分为 A、B、C、D、E 5 组，每组 6 只；制备直径 1 cm 双侧胫骨缺损模型后，A 组不植入任何材料，B～E 组分别于骨缺损处植入支架 3、支架 4、支架 1、支架 2。术后 4、8、12 周大体观察骨缺损修复效果；行 HE、Masson 染色以及成骨特异性转录因子 RUNX2、成骨相关转录因子 Osterix（OSX）、ColⅠ、骨桥蛋白（osteopontin，OPN）抗体的免疫组织化学染色，观察不同支架材料修复骨缺损效果。结果扫描电镜观察示，支架 2 为多孔结构，交联前结构疏松，交联后结构致密；表面接触角检测示支架为疏水性材料，且支架 2 疏水性比支架 4 更高；药物体外缓释效果显示支架 2 上药物能微量长效释放。动物体内植入实验中，大体观察示术后 4、12 周 D、E 组缺损明显小于 A、B、C 组。HE 和 Masson 染色示，术后 4 周 A 组缺损区域充满大量结缔组织，B、C 组可见大量纤维组织，D、E 组可见少量新生骨；术后 8 周各组新生骨比 4 周时增加；术后 12 周 A 组缺损区域仍以纤维组织为主，B、C 组可见少量新生骨组织，D、E 组可见大量新生骨组织，尤以 E 组明显，且大部分支架降解。免疫组织化学染色示，术后 4 周，D、E 组新生组织中 RUNX2 和 OSX 表达明显高于其余各组，术后 8、12 周 RUNX2 表达与 4 周相比表达下降；术后 8、12 周与 4 周相比，各组 ColⅠ、OPN 表达均增加，且 D、E 组新生组织中 ColⅠ、OPN 表达明显高于其余各组。结论ICA/ATP/Col Ⅰ/PCL 复合支架具有良好的孔隙率和生物相容性，并具有促成骨作用，可达到良好骨再生和修复效果。     

冰片对黄芪甲苷和三七总皂苷配伍有效成分在脑缺血/再灌注模型大鼠脑组织分布的影响

目的探讨冰片是否具有促进黄芪甲苷(AST IV)和三七总皂苷(PNS)配伍时主要有效成分透过大脑中动脉栓塞(MCAO)再灌注模型大鼠血脑屏障的作用。方法大鼠随机分为假手术组、模型组、冰片组、AST IV组、PNS组、AST IV+PNS组、冰片+AST IV+PNS组,制备MCAO再灌注大鼠模型,以液相色谱-质谱联用法(LC-MS/MS)测定大鼠患侧与健侧大脑皮层、小脑中AST IV和PNS有效成分(人参皂苷Rg1、Rb1和三七皂苷R1)的含量。结果 AST IV无论是单用还是与PNS、冰片配伍,其口服后主要分布在大脑皮层,尤其是患侧大脑皮层。冰片+AST IV+PNS能使患侧与健侧大脑皮层中AST IV含量显著增加。PNS单用,其有效成分人参皂苷Rg1、Rb1和三七皂苷R1主要分布在患侧小脑。冰片+AST IV+PNS能使患侧大脑皮层中人参皂苷Rb1含量显著增加,使健侧和患侧大脑皮层中人参皂苷Rg1含量增加,使大脑皮层尤其是患侧大脑皮层及小脑中三七皂苷R1含量增加。结论大鼠脑缺血再灌注后,AST IV与PNS的有效成分人参皂苷Rb1、Rg1及三七皂苷R1在大脑皮层和小脑均有一定的分布。AST IV单用时,AST IV主要分布在大脑皮层;PNS单用时,人参皂苷Rb1、Rg1及三七皂苷R1主要分布在小脑。冰片与AST IV、PNS合用后,能促进AST IV及人参皂苷Rb1、Rg1及三七皂苷R1向大脑皮层富集,尤其是向缺血再灌注侧大脑皮层富集;而且能不同程度地促进AST IV,人参皂苷Rb1、Rg1及三七皂苷R1在大脑皮层的吸收,尤其是在患侧大脑皮层的吸收。     

Optimal Respiratory Syncytial Virus intervention programmes using Nirsevimab in England and Wales

IntroductionRespiratory Syncytial Virus (RSV) is a major cause of acute lower respiratory tract infections (ALRI) in infants. There are no licensed vaccines and only one monoclonal antibody available to protect infants from disease. A new and potentially longer-lasting monoclonal antibody, Nirsevimab, showed promising results in phase IIb/III trials. We evaluate the cost-effectiveness of Nirsevimab intervention programmes in England and Wales.MethodsWe used a dynamic model for RSV transmission, calibrated to data from England and Wales. We considered a suite of potential Nirsevimab programmes, including administration to all neonates (year-round); only neonates born during the RSV season (seasonal); or neonates born during the RSV season plus infants less than six months old before the start of the RSV season (seasonal + catch-up).ResultsIf administered seasonally to all infants at birth, we found that Nirsevimab would have to be priced at £63 or less per dose for at least 50% certainty that it could cost-effectively replace the current Palivizumab programme, using an ICER threshold of £20,000/QALY. An extended seasonal programme which includes a pre-season catch-up becomes the optimal strategy at a purchasing price of £32/dose or less for at least 50% certainty. At a purchasing price per dose of £5-32, the annual implementation costs of a seasonal programme could be as high as £2 million before a switch to a year-round strategy would be optimal.DiscussionNirsevimab has the potential to be cost-effective in England and Wales not only for use in high-risk infants.     

Midterm Outcomes of Angioplasty for Pediatric Renovascular Hypertension

PurposeTo evaluate the midterm outcomes of percutaneous transluminal renal angioplasty (PTRA) for pediatric renovascular hypertension (RVH).Materials and MethodsThe clinical data of patients who underwent PTRA for RVH in the authors’ hospital from 2012 to 2019 were retrospectively analyzed. Postprocedural blood pressure, glomerular filtration rate (GFR) of the affected kidney, restenosis, and complications were closely monitored.ResultsPTRA was performed in a total of 30 children (20 boys and 10 girls), with a mean age of 7.3 years ± 0.7 (range, 40 days to 13.9 years) and a mean weight of 25.0 kg ± 2.3 (range, 3.4–53 kg). The median follow-up period was 26.5 months (range, 1 month to 7.5 years). Technical success was achieved in 26 (86.7%) of the 30 patients. Restenosis developed in 3 patients (10.0%). Only 1 patient underwent stent implantation, and the stent fractured 8 months later, requiring further intervention. There were no other complications. In terms of clinical benefit of blood pressure control after the initial PTRA procedure, 15 patients (50%) were cured and 7 patients (23.3%) showed improvement. There was no significant difference in the etiology, lesion location, and lesion length between patients with clinical benefit and failure (P = .06, P = .202, and P = .06, respectively). GFR of the affected kidney was significantly improved from 19.9 mL/min ± 11.2 to 38.1 mL/min ± 11.9 at the 6-month follow-up after PTRA (P < .001).ConclusionsThe overall results of PTRA for pediatric RVH caused by different etiologies are promising. PTRA not only provided a clinical benefit of blood pressure control in 73.3% of the patients but also significantly improved the function of the affected kidney.