Clinical application of metagenomic next‐generation sequencing technology in the diagnosis and treatment of pulmonary infection pathogens: A prospective single‐center study of 138 patients

IntroductionRapid and accurate pathogen identification is essential for the treatment of pneumonia. Metagenomic next‐generation sequencing (mNGS) is a newly developed technology to obtain microbial nucleic acid sequence information quickly, efficiently, and without bias.MethodsWe performed shotgun metagenomic next‐generation sequencing (mNGS) of bronchoalveolar lavage fluid (BALF) for pathogen identification in pneumonia in a prospective study with 138 patients from a single center. We compared the results of mNGS with standard methods including culture, staining, and targeted PCR and evaluated the clinical applicability of mNGS.ResultsMost of the patients (128/138, 92.75%) were cured or improved. One patient (1/138, 0.72%) died because of acute gastrointestinal bleeding, and 9 patients (9/138, 6.52%) showed no improvement. mNGS identified more bacteria (53 versus 27), fewer fungi (8 versus 31), and more viruses (16 versus 1) than standard methods. In total, treatment in 34 out of 138 cases (24.64%) was adjusted and optimized because of mNGS results. Positive mNGS results contributed to a definitive diagnosis in 23 cases (16.67%), which helped guide treatment decision by either adjusting the antibiotics without de‐escalation or continuing the empirical treatment. mNGS also confirmed no active infection in 11 cases (7.97%) allowed for antibiotic de‐escalation.ConclusionThis prospective clinical study evaluated the clinical utility of mNGS for the diagnosis of pneumonia and showed that mNGS of BALF provides valuable information for effective treatment.     

Development and Primary Application of an Indirect ELISA Based on Rep Protein to Analyze Antibodies against Porcine Cocirvirus-like Virus (PCLV)

Porcine circovirus-like virus (PCLV) is a member of circovirus that contains a single-strand DNA genome, which may be one of the pathogens that causes diarrheal symptoms in pigs. The Rep protein encoded by the genome of PCLV may be responsible for viral genome replication. The development of serological detection methods for PCLV is of great necessity for clinical diagnosis, as well as epidemiological investigations. Therefore, this study attempted to build an indirect enzyme-linked immunosorbent assay (ELISA) to examine antibodies against PCLV based on the His-tagged recombinant Rep protein. Full-length PCLV Rep protein was induced and expressed in E. coli and was purified as an antigen to establish an ELISA detection kit. The purified Rep protein was used to inject into mice to produce specific antibodies. There was no cross-reaction of Rep-based ELISA with antisera against other porcine viruses. The intra-assay and inter-assay coefficient variations (CVs) were 0.644–8.211% and 0.859–7.246%, respectively, indicating good repeatability. The non-cross-reaction with TGEV, PRRSV and PCV2 testing showed high sensitivity and high specificity for this ELISA assay. A total of 1593 serum samples collected from different pig farms in Jiangxi Province were tested for anti-PCLV Rep antibodies, and 284 (17.83%) of the 1593 samples were Rep antibody positive. Altogether, the indirect ELISA detection tool developed in this study could be applied to examine serum of PCLV antibodies with good repeatability, high sensitivity and high specificity. In addition, field sample detection results suggested that the PCLV antibody has a low prevalence in pig populations in Jiangxi Province of China.     

正常中国汉人红细胞中儿茶酚氧位甲基转移酶的活性测定

目的测定正常中国人群红细胞儿茶酚氧位甲基转移酶(COMT)活性浓度值,为研究COMT活性变化在疾病诊断中的应用提供依据。方法采用高效液相色谱法测定279名正常健康人(其中男性134例,女性145例)红细胞中COMT的活性浓度,用SPSS统计软件对性别间测定结果进行检验比较。结果男性和女性红细胞中COMT活性浓度分别为:(16.5±7.4)、(15.5±5.3)nmol.mL RBC-1.h-1。活性频率分布男性在6.0~37.4nmol.mL RBC-1.h-1,女性为5.4~37.2 nmol.mL RBC-1.h-1,男女之间COMT活性浓度的差异无统计学意义(P>0.05)。结论本法灵敏,简便,准确,重复性好,且测定结果与国外一些报道一致,适用于临床应用测定研究。本结果可作为中国健康人红细胞COMT活性正常值及今后研究的重要参考数据。     

Cell-penetrating peptide-doxorubicin conjugate loaded NGR-modified nanobubbles for ultrasound triggered drug delivery

A new drug-targeting system for CD13⁺ tumors has been developed, based on ultrasound-sensitive nanobubbles (NBs) and cell-permeable peptides (CPPs). Here, the CPP-doxorubicin conjugate (CPP-DOX) was entrapped in the asparagine–glycine–arginine (NGR) peptide modified NB (CPP-DOX/NGR-NB) and the penetration of CPP-DOX was temporally masked; local ultrasound stimulation could trigger the CPP-DOX release from NB and activate its penetration. The CPP-DOX/NGR-NBs had particle sizes of about 200?nm and drug entrapment efficiency larger than 90%. In vitro release results showed that over 85% of the encapsulated DOX or CPP-DOX would release from NBs in the presence of ultrasound, while less than 1.5% of that (30?min) without ultrasound. Cell experiments showed the higher cellular CPP-DOX uptake of CPP-DOX/NGR-NB among the various NB formulations in Human fibrosarcoma cells (HT-1080, CD13⁺). The CPP-DOX/NGR-NB with ultrasound treatment exhibited an increased cytotoxic activity than the one without ultrasound. In nude mice xenograft of HT-1080 cells, CPP-DOX/NGR-NB with ultrasound showed a higher tumor inhibition effect (3.1% of T/C%, day 24), longer median survival time (50 days) and excellent body safety compared with the normal DOX injection group. These results indicate that the constructed vesicle would be a promising drug delivery system for specific cancer treatment.     

Investigation of Potential Use of Soybean Protein Isolate–Chinese Bayberry Tannin Extract Cross-Linked Films in Packaging Applications

The possibility of using commercial bayberry tannin (BT) from a Chinese source as a cross-linker and functional additive to develop soybean protein isolate (SPI)-based films was explored in this study by using the solvent casting method. In particular, the impacts of BT loading on the tensile strength, microstructure, thermal stability, water resistance and antioxidant capacity were fully investigated. The results reveal that SPI incorporated with BT yielded a phenolic–protein hybrid whose relevant films exhibited an improvement in tensile strength of around two times greater compared with native SPI as a result of the formed interactions and covalent cross-links, which could be proven using FTIR spectroscopy. The introduction of BT also led to the compact microstructure of SPI–BT films and enhanced the thermal stability, while the water vapor permeability was reduced compared with the control SPI film, especially at high loading content of tannin. Additionally, the use of BT significantly promoted the antioxidant capacity of the SPI-based films according to DPPH radical scavenging assay results. On this basis, Chinese bayberry tannin is considered a promising natural cross-linker and multifunctional additive that can be dedicated to developing protein-derived films with antioxidant activity for food packaging applications.     

促红细胞生成素与促红细胞生成素受体在大鼠脱离视网膜中的表达

目的 观察促红细胞生成素（EPO）和促红细胞生成素受体（EPOR）在大鼠脱离视网膜中的表达情况。方法48只雄性SD大鼠,随机分为正常对照组、视网膜脱离（RD）后1、3、6、12、24、48、72 h组,每组6只大鼠12只眼。视网膜下腔单次注射1.4%透明质酸钠致上半侧视网膜隆起建立RD模型,采用逆转录聚合酶链式反应（RT-PCR）和蛋白免疫印迹（Western blot）测定不同时间点EPO和EPOR的mRNA和蛋白水平的表达情况,同时采用免疫组织化学方法观察EPO和EPOR在视网膜中定位表达的情况。结果 EPO和EPOR的mRNA表达水 平在RD后均上调,均于RD后48 h达到高峰,分别于RD后6、12 h显著高于正常对照组,差异具有统计学意义（P＜0.05）;同样,EPO和EPOR的蛋白表达水平也在RD后均增高并在RD后48 h达到高峰,均于RD后3 h显著高于正常对照组,差异具有统计学意义（P＜0.05）。免疫组织化学结果显示正常视网膜自神经节细胞层至光感受器细胞的内外节均有EPO弱表达,RD后48 h视网膜相应部位呈强阳性表达;正常视网膜自神经节细胞层至光感受器细胞的内节均有EPOR表达,RD后48 h视网膜相应部位呈强阳性着染。结论 RD后大鼠视网膜EPO和EPOR表达均逐渐增强,48 h达到高峰;大鼠神经视网膜大部分层次能表达EPO和EPOR。     

鼓室硬化和非鼓室硬化中耳炎抗氧化能力的差异性研究

目的 了解鼓室硬化和非鼓室硬化中耳炎患者在抗氧化物质超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalnse,CAT)活性及过氧化反应产物丙二醛(malondialdehyde,MDA)浓度方面的差异,以进一步探讨鼓室硬化形成的机制.方法 回顾分析56例(56耳)中耳炎、中耳炎后遗症手术患者资料,分成两组:鼓室硬化组26耳,非鼓室硬化组30耳.测定所有患者血浆、红细胞中SOD、CAT活力及血浆、中耳组织中MDA浓度.结果 分析用秩和检验及相关分析.结果 两组血浆中MDA含量差异无统计学意义.鼓室硬化组中耳组织中MDA含量较高,差异有统计学意义(P<0.05).两组红细胞中SOD和CAT活力差异无统计学意义.鼓室硬化组血浆中的SOD和CAT活力较非鼓室硬化组低.差异有统计学意义.鼓室硬化组术前的听力平均值与其各检测项目所测得的MDA、SOD、CAT值均不存在相关(P>0.05).结论 鼓室硬化患者可能存在局部氧自由基浓度增高和抗氧化物质活性降低的情况.局部氧自由基浓度增高,血液抗氧化物质无法进入中耳组织,可能是形成鼓室硬化的病理基础.     

Safety and efficacy of anti-EGFR monoclonal antibody (SCT200) as second-line therapy in advanced esophageal squamous cell carcinoma

Objective: The mainstay treatment of esophageal squamous cell carcinoma(ESCC) involves chemotherapy and immunotherapy. However, alternative therapies are required for patients who are refractory or intolerant to existing therapies.Methods: In this single-arm, multicenter, open-label phase Ib study, 30 patients received an intravenous infusion of SCT200, an antiepidermal growth factor receptor(EGFR) monoclonal antibody, 6.0 mg/kg once a week for 6 weeks, followed by 8.0 mg/kg once every 2 weeks u...     

Construction of developmentally inspired periosteum-like tissue for bone regeneration

The periosteum, a highly vascularized thin tissue, has excellent osteogenic and bone regenerative abilities. The generation of periosteum-mimicking tissue has become a novel strategy for bone defect repair and regeneration, especially in critical-sized bone defects caused by trauma and bone tumor resection. Here, we utilized a bone morphogenetic protein-2(BMP-2)-loaded scaffold to create periosteum-like tissue(PT) in vivo, mimicking the mesenchymal condensation during native long bone developmen...     

植物雌激素与性激素失调导致的干眼症

植物雌激素是一类天然的结构及生物活性均类似于雌激素的非甾体类化合物，可与雌激素受体结合，对内源性雌激素起双向调节作用。多种原因引起的性激素失调可以导致干眼症。作者认为植物雌激素因其特有的优势可以治疗此类干眼症，值得进一步研究。