一款用于快速检测包括SARS-CoV-2在内的19种常见呼吸道病毒的高通量、多指标等温扩增平台

快速、准确地诊断以及及时对感染严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的患者进行隔离被认为是抗击新冠病毒肺炎(COVID-19)大流行的最有效的措施。本文介绍了一款高通量、多指标的等温扩增检测平台(RTisochip^TM-W),该平台使用离心式微流控芯片,在90min内即可完成一次运行,从16个样本中检测出包括SARS-CoV-2在内的19种常见呼吸道病毒。通过RTisochip^TM-W系统检测的所有病毒的检出限均等于或小于50拷贝·μL^-1,与常规逆转录聚合酶链反应(RT-PCR)灵敏度相当。该系统具有重复性好、鲁棒性强和特异性高的优点。最后,使用RTisochip^TM-W系统对201例临床前样本、14例COVID-19阳性临床样本、25例临床确诊样本以及614例来自呼吸道感染或疑似感染的患者的临床样本进行了测试。测试结果与临床诊断结果吻合良好,也反映出呼吸道传染病的流行特征。在对SARS-CoV-2的诊断上,RTisochip^TM-W系统与参考试剂盒的一致性达到98.15%。基于大量的测试结果,我们相信RTisochip^TM-W系统是一款抗击COV ID-19大流行的强有力工具。

The Current State and Progress of Mpox Vaccine Research

On July 23,2022,the World Health Organization(WHO)declared the monkeypox(mpox)outbreak a“Public Health Emergency of International Concern.”Since 2022,outbreaks of mpox in many countries around the world have primarily resulted in fatalities among immunocompromised individuals,such as untreated HIV/AIDS patients.Since the eradication of smallpox was declared by the WHO in 1980,the global vaccination against smallpox has been gradually discontinued.China also stopped routine smallpox vaccination in 1981.The protective effect of the smallpox vaccine has decreased over time due to aging and declining immunity in those who were vaccinated.For individuals,timely vaccination against smallpox is an effective means of protection against mpox.However,due to safety concerns with the smallpox vaccine and the limitations of current mpox vaccines,there is no vaccine that is safe,effective,and has low side effects applied in clinical settings.This article provides a comprehensive review of the development of mpox virus(MPXV)vaccines,their application in special populations,and the current state of vaccine research,considering the etiology,transmission,and prevention of the MPXV.Vaccination,as an effective method of epidemic prevention,can provide long-term immune protection and effectively reduce the severity of infection.However,as there is no licensed specific MPXV vaccine available globally,the vaccines currently used for mpox prevention are mostly smallpox vaccines.These smallpox vaccines can offer some degree of protection against mpox by activating cross-protection in the body.

三维生物打印抗菌型水凝胶对大鼠全层皮肤缺损创面的作用

目的探讨三维生物打印负载纳米银的甲基丙烯酸酐化明胶(GelMA)水凝胶对大鼠全层皮肤缺损创面的作用。方法采用实验研究方法。采用扫描电子显微镜观察不同质量浓度的纳米银溶液中的纳米银颗粒的形态、粒径、分布和含不同终质量分数GelMA的含银GelMA水凝胶的孔隙结构,并计算孔径大小。处理1、3、7、14 d,采用质谱仪检测含终质量分数15%GelMA和终质量浓度10 mg/L纳米银的水凝胶的纳米银释放浓度。培养24 h,检测含终质量浓度0(无纳米银)、25、50、100 mg/L纳米银的GelMA水凝胶对金黄色葡萄球菌和大肠埃希菌的抑菌圈直径。取浙江大学医学院附属第二医院泌尿外科2020年7月收治的1名健康5岁男童包皮环切术后废弃包皮和该院整形外科2020年7月收治的1名健康23岁女性抽脂手术后废弃脂肪,采用酶解法分别提取成纤维细胞(Fb)和脂肪干细胞(ASC)。将Fb分为仅有培养基的空白对照组和另加入含相应终质量浓度纳米银溶液的2 mg/L纳米银组、5 mg/L纳米银组、10 mg/L纳米银组、25 mg/L纳米银组、50 mg/L纳米银组,培养48 h,采用细胞计数试剂盒8检测Fb增殖活性。将Fb分为进行相应处理的0 mg/L含银GelMA水凝胶组、10 mg/L含银GelMA水凝胶组、50 mg/L含银GelMA水凝胶组、100 mg/L含银GelMA水凝胶组,于培养1、3、7 d,同前检测Fb增殖活性。将ASC与GelMA水凝胶混合种植,分为三维生物打印组和非打印组,于培养1、3、7 d,同前检测ASC增殖活性并行活/死细胞荧光染色观察ASC生长情况。以上实验中样本数均为3。于18只雄性4~6周龄SD大鼠背部各制作4个全层皮肤缺损创面,分别设为单纯水凝胶组、水凝胶/纳米银组、水凝胶支架/纳米银组、水凝胶支架/纳米银/ASC组并移植相应支架。分别于伤后4、7、14、21 d,行大体观察并计算创面愈合率(样本数为6);于伤后7、14 d,对创面行苏木精-伊红染色观察组织病理学改变(样本数为6);于伤后21 d,对创面行Masson染色观察胶原排列情况(样本数为3)。对数据行单因素方差分析、重复测量方差分析、Bonferroni校正、独立样本t检验。结果不同质量浓度的纳米银溶液中的纳米银颗粒均呈圆形,散在分布,粒径均匀。含不同终质量分数GelMA的含银GelMA水凝胶都呈现大小不一且相互连通的孔隙结构。含终质量分数10%GelMA的含银GelMA水凝胶的孔径明显大于含终质量分数15%和20%GelMA的含银GelMA水凝胶(P值均<0.05)。处理1、3、7 d,含银GelMA水凝胶的体外纳米银释放浓度的变化趋势相对平缓;处理14 d,其体外纳米银释放浓度迅速增加。培养24 h,含0、25、50、100 mg/L纳米银的GelMA水凝胶对金黄色葡萄球菌和大肠埃希菌的抑菌圈直径分别为0、0、0.7、2.1 mm和0、1.4、3.2、3.3 mm。培养48 h,2 mg/L纳米银组、5 mg/L纳米银组Fb的增殖活性均明显高于空白对照组(P<0.05),10 mg/L纳米银组、25 mg/L纳米银组、50 mg/L纳米银组Fb的增殖活性均明显低于空白对照组(P<0.05)。与0 mg/L含银GelMA水凝胶组相比,培养1 d,50 mg/L含银GelMA水凝胶组、100 mg/L含银GelMA水凝胶组Fb的增殖活性均明显降低(P<0.05);培养3 d,50 mg/L含银GelMA水凝胶组Fb的增殖活性明显升高(P<0.05),100 mg/L含银GelMA水凝胶组Fb的增殖活性明显降低(P<0.05);培养7 d,100 mg/L含银GelMA水凝胶组Fb的增殖活性明显降低(P<0.05)。培养1 d,三维生物打印组的ASC增殖活性与非打印组比较,差异无统计学意义(P>0.05);培养3、7 d,三维生物打印组的ASC增殖活性均明显高于非打印组(t值分别为21.50、12.95,P<0.05)。培养1 d,三维生物打印组死亡ASC数略多于非打印组。培养3、5 d,三维生物打印组和非打印组中的绝大多数ASC为活细胞。伤后4 d,单纯水凝胶组和水凝胶/纳米银组大鼠创面渗液较多,水凝胶支架/纳米银组和水凝胶支架/纳米银/ASC组大鼠创面干燥且未见明显感染迹象。伤后7 d,单纯水凝胶组和水凝胶/纳米银组大鼠创面仍有少量渗液,水凝胶支架/纳米银组和水凝胶支架/纳米银/ASC组大鼠创面干燥结痂。伤后14 d,4组大鼠创面上的水凝胶均脱落。伤后21 d,仅单纯水凝胶组仍有少量创面未愈合。伤后4、7 d,水凝胶支架/纳米银/ASC组大鼠创面愈合率均明显高于其他3组(P<0.05)。伤后14 d,水凝胶支架/纳米银/ASC组大鼠创面愈合率明显高于水凝胶/纳米银组和单纯水凝胶组(P值均<0.05)。伤后21 d,单纯水凝胶组大鼠创面愈合率明显低于水凝胶支架/纳米银/ASC组(P<0.05)。伤后7 d,4组大鼠创面上的水凝胶均保持在位;伤后14 d,单纯水凝胶组大鼠的水凝胶已与创面脱离,而其余3组仍有部分水凝胶存在于创面新生组织中。伤后21 d,单纯水凝胶组大鼠创面胶原排列无序,而水凝胶/纳米银组、水凝胶支架/纳米银组、水凝胶支架/纳米银/ASC组大鼠创面胶原排列相对有序。结论含银GelMA水凝胶具有良好的生物相容性及抗菌性能,其三维生物打印的双层结构能更好地与大鼠全层皮肤缺损创面新生组织相融合并促进创面愈合。

T cell-depleting nanoparticles ameliorate bone loss by reducing activated T cells and regulating the Treg/Th17 balance

Estrogen deficiency is one of the most frequent causes of osteoporosis in postmenopausal women.Under chronic inflammatory conditions caused by estrogen deficiency,activated T cells contribute to elevated levels of proinflammatory cytokines,impaired osteogenic differentiation capabilities of bone marrow mesenchymal stem cells(BMMSCs),and disturbed regulatory T cell(Treg)/Th17 cell balance.However,therapeutic strategies that re-establish immune homeostasis in this disorder have not been well developed.Here,we produced T cell-depleting nanoparticles(TDNs)that ameliorated the osteopenia phenotype and rescued the osteogenic deficiency of BMMSCs in ovariectomized(OVX)mice.TDNs consist of monocyte chemotactic protein-1(MCP-1)-encapsulated mesoporous silica nanoparticles as the core and Fas-ligand(FasL)as the corona.We showed that the delicate design of the TDNs enables rapid release of MCP-1 to recruit activated T cells and then induces their apoptosis through the conjugated FasL both in vitro and in vivo.Apoptotic signals recognized by macrophages help skew the Treg/Th17 cell balance and create an immune tolerant state,further attenuating the osteogenic deficiency of BMMSCs and the osteopenia phenotype.Mechanistically,we found that the therapeutic effects of TDNs were partially mediated by apoptotic T cell-derived extracellular vesicles(ApoEVs),which promoted macrophage transformation towards the M2 phenotype.These findings demonstrate that TDNs may represent a promising strategy for treating osteoporosis and other immune disorders.

灭活疫苗或腺病毒载体疫苗初次免疫后不同疫苗加强针中和SARS-CoV-2 Omicron变异株的效果

SARS-CoV-2 Omicron变异株已在全球范围内广泛传播.包括中国在内的许多国家广泛接种了SARS-CoV-2灭活疫苗(IAV)、重组蛋白疫苗(PRV)和腺病毒载体疫苗(Ad V),以达到群体免疫效果.该研究系统评价了完成不同疫苗全程接种后以及接种不同疫苗加强针后的341份人血清对原始毒株和Omicron变异株的交叉中和活性.研究发现,所有类型疫苗诱导的针对Omicron变异株BA.1的中和抗体水平都明显低于原始毒株.初免疫IAV和Ad V后,分别接种Ad V和PRV的异源加强针可以最大程度地提高血清对Omicron中和活性.此外,该研究使用m RNA疫苗在小鼠模型中评估了初次免疫编码Spike蛋白的m RNA疫苗后,加强免疫编码原始毒株或者不同突变株RBD蛋白的m RNA疫苗对Omicron中和效果.结果发现,与原始毒株加强针相比,只有Omicron加强针疫苗显著地提升了Omicron的中和抗体水平.该研究对各种疫苗加强针的系统性评价可为接下来对抗Omicron变异株的免疫接种提供参考.

Curcumin-incorporated 3D bioprinting gelatin methacryloyl hydrogel reduces reactive oxygen species-induced adipose-derived stem cell apoptosis and improves implanting survival in diabetic wounds

Background:Gelatin methacryloyl(GelMA)hydrogels loaded with stem cells have proved to be an effective clinical treatment for wound healing.Advanced glycation end product(AGE),interacting with its particular receptor(AGER),gives rise to reactive oxygen species(ROS)and apoptosis.Curcumin(Cur)has excellent antioxidant activity and regulates intracellular ROS production and apoptosis.In this study,we developed a Cur-incorporated 3D-printed GelMA to insert into adiposederived stem cells(ADSCs)and applied it to diabetic wounds.Methods:GelMA hydrogels with Cur were fabricated and their in vitro effects on ADSCs were investigated.We used structural characterization,western blot,ROS and apoptosis assay to evaluate the antioxidant and anti-apoptotic activity,and assessed the wound healing effects to investigate the mechanism underlying regulation of apoptosis by Cur via the AGE/AGER/nuclear factor-κB(NF-κB)p65 pathway.Results:A 10%GelMA scaffold exhibited appropriate mechanical properties and biocompatibility for ADSCs.The circular mesh structure demonstrated printability of 10%GelMA and Cur-GelMA bioinks.The incorporation of Cur into the 10%GelMA hydrogel showed an inhibitory effect on AGEs/AGER/NF-κB p65-induced ROS generation and ADSC apoptosis.Furthermore,Cur-GelMA scaffold promoted cell survival and expedited in vivo diabetic wound healing.Conclusions:The incorporation of Cur improved the antioxidant activity of 3D-printed GelMA hydrogel and mitigated AGE/AGER/p65 axis-induced ROS and apoptosis in ADSCs.The effects of scaffolds on wound healing suggested that Cur/GelMA-ADSC hydrogel could be an effective biological material for accelerating wound healing.

Evaluation of the immunomodulatory effects of anti-COVID-19 TCM formulae by multiple virus-related pathways

Dear Editor,The coronavirus infectious disease 2019(COVID-19)outbreak is seriously endangering human health.Most patients with severe COVID-19 are characterized by sustained cytokine production and hyper-inflammation,which is known as cytokine storm syndrome.1-3 Elaborating the anti-inflammatory response is crucial to these patients,and interieukin-6(IL6)inhibitors and steroids have been recommended in clinical practice.1 However,after the cytokine storm phase,the host immune response to sepsis may develop into a protracted immunosuppressive phase.

Quantification of SARS-CoV-2 antigen levels in the blood of patients with COVID-19

Dear Editor,Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has caused the global outbreak of coronavirus disease 2019(COVID-19).By far,more than 35 million people had been infected by SARS-CoV-2,resulting in more than 1 million deaths globally.It is well recognized that SARSCoV-2 preferentially attacks pulmonary epithelial cells,leading to acute respiratory distress syndrome(ARDS).

High-dimensional immune profiling by mass cytometry revealed immunosuppression and dysfunction of immunity in COVID-19 patients

The outbreak of coronavirus disease 2019(COVID-19)caused by the new virus SARS-CoV-2 has been announced as a public health emergency of international concern.1–3 The clinical features of patients with COVID-19 range from common fever and cough to other rare symptoms,such as diarrhea and nausea.This disease can progress quickly,and 2–3%of patients die within a short time,which is generally due to multiple organ failure.4–7 Clinically,COVID-19 patients are classified into mild,moderate,severe,and critical cases.5–7 The immune response against SARS-CoV-2 is probably linked to the severity of disease.Recently,Zheng et al.8 showed that elevated levels of T-cell exhaustion and reduced functional diversity of T cells in peripheral blood may predict severe progression in COVID-19 patients;however,a more comprehensive understanding of the pathology of SARS-CoV-2 infection remains to be delineated.Here,we profiled immune cellular components using mass cytometry(CyTOF)to analyze the peripheral blood mononuclear cells(PBMCs)from patients with differences in disease progression by comparing with the PBMCs from healthy donors(HDs).

The 200-hPa Wind Perturbation in the Subtropical Westerly over East Asia Related to Medium-Range Forecast of Summer Rainfall in China

The 200-hPa wind perturbation(WP) in the subtropical westerly over East Asia(SWEA) has seldom been examined in previous studies, especially in connection with forecast of the summer rainfall in China. Based on the daily NCEP/NCAR reanalysis data and precipitation observations in China from 1 June to 31 August of 1960-2015, this study first systematically analyzes the spatiotemporal distribution features of the 200-hPa WP in the SWEA on different scales, especially during the Meiyu season in the Yangtze-Huaihe region and during the rainy period in North China, by using spectral decomposition and period analysis. It is found that in the 56-yr mean fields, the 200-hPa WP in the SWEA is collocated with the East Asian subtropical jet(EASJ), with the centers of the two systems coincidentally overlapped. The WP filed in the subtropical westerly mainly comprises planetary-and synoptic-scale waves. The quasi-stationary planetary-scale wave seems to determine the shape and intensity of the WP in the SWEA, while the synoptic-scale wave is closely related to the local central intensity of the WP. The daily evolution of the 56-yr mean fields shows that, following the northward(southward) movement of SWEA from 1 June to 31 August, the planetaryscale WP in the SWEA becomes gradually weakened(intensified) whereas the synoptic-scale WP is slightly intensified(weakened).The results also reveal that during the Meiyu season in the Yangtze-Huaihe region, the WP in the SWEA moves northward slowly around 37°-39°N, demonstrating a quasi-biweekly oscillation in its geographic location, which is largely attributed to the strong(weak) planetary-scale(synoptic-scale) component. On the contrast, in the summer rainy season over North China, the WP in the SWEA further strives northward beyond 40°N, showing both quasi-biweekly and weekly oscillations in its position; meanwhile, the planetary-scale wave in the SWEA becomes weakened whereas the synoptic-scale wave is enhanced. These salient variational features of the WP in the SWEA and its scaledependent components may be useful for the medium-range forecast of the rain belt migration in eastern China.

Redox-responsive micelles integrating catalytic nanomedicine and selective chemotherapy for effective tumor treatment

Chemotherapy is one of the most conventional modalities for cancer therapy.However,the high multidrug resistance of tumor cells still limited the clinical application of current chemotherapy.Considering the ability of nitric oxide(NO) to modulate potent P-glycoprotein to inhibit multi-drug resistance,a synergistic methodology combining chemotherapy and sustained NO generation is an ideal way to further promote the chemotherapy.Herein,a multi-functional micelle with tumor-selective chemotherapy driven by redox-triggered doxorubicin(DOX) release and drug resistance inhibition based on intracellular NO generation was fabricated for effective tumor treatment.The micelle consists of DOX as core,arginine/glucose oxidase(Arg/GOx) as shell and redox-responsive disulfide bond as a linker,which is denoted as micelle-DOX-Arg-GOx.The Arg serves as the biological precursor of nitric oxide for inhibition of multi-drug resistance to promote chemotherapy and GOx catalyzes glucose to produce hydrogen peroxide(H_(2) O_(2)) for increasing the generation of NO.Moreover,the glucose supply could be simultaneously blocked by the catalytic process,which further enhanced therapeutic efficiency.This micelle requests a tumor-specific microenvironment(a considerable amount of GSH) to perform synergistic therapeutics including chemotherapy,starvation therapy(catalytic medicine),and gas therapy for tumor treatment,which resulted in significant cytotoxicity to tumor tissue.

Forced Decadal Changes in Summer Precipitation Characteristics over China:The Roles of Greenhouse Gases and Anthropogenic Aerosols

We investigated the decadal changes in the different types of summer mean precipitation over China across the mid-1990 s based on observational datasets.The spatial variations in the observed decadal changes were estimated by comparing the present day(PD)time period of 1994–2011 with an earlier period of 1964–1981.The summer total precipitation increased in southern China and decreased in northern China from the early period to the PD.The increases of precipitation in southern China were due to increases in the frequency of heavy and moderate rainfall,whereas the decreases over northern China were mainly due to decreases in the frequency of moderate and light rainfall.Based on a set of numerical experiments using an atmospheric general circulation model coupled with a multilevel mixed-layer ocean model,we found that the increase of precipitation frequency forced by greenhouse gases is the main reason of increasing precipitation over southern and northeastern China,while the decrease of frequency caused by anthropogenic aerosol(AA)induces the decreasing precipitation over northern China.The water vapor flux convergence and water vapor flux strengthen in southern China and northeastern China by anthropogenic greenhouse gases.This distribution is also conducive to precipitation in most of southern China and northeastern China.Under the control of weakened southwesterly winds and 850-h Pa divergence,precipitation decreases over northern and southwestern China by AA.

Single-cell RNA sequencing-based immunological feature analysis of a COVID-19 patient with recurrent positive SARS-CoV-2 RNA

To the Editor:Since the outbreak in December 2019,severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has infected>2.4 billion people worldwide,resulting in>4.9 million deaths.To understand the pathogenesis of SARS-CoV-2 infection,single-cell RNA-sequencing(scRNA-seq)profiles of peripheral blood mononuclear cells(PBMCs)have been constructed in coronavirus disease-2019(COVID-19)patients at different disease stages.