Self-confocal NIR-II fluorescence microscopy for multifunctional in vivo imaging

Fluorescence imaging in the second near-infrared window(NIR-II,900–1880 nm)with less scattering background in biological tissues has been combined with the confocal microscopic system for achieving deep in vivo imaging with high spatial resolution.However,the traditional NIR-IIfluorescence confocal microscope with separate excitation focus and detection pinhole makes it possess low confocal e±ciency,as well as di±cultly to adjust.Two types of upgraded NIR-IIfluorescence confocal microscopes,sharing the same pinhole by excitation and emission focus,leading to higher confocal e±ciency,are built in this work.One type is-ber-pinhole-based confocal microscope applicable to CW laser excitation.It is constructed forfluorescence intensity imaging with large depth,high stabilization and low cost,which could replace multiphotonfluorescence microscopy in some applications(e.g.,cerebrovascular and hepatocellular imaging).The other type is air-pinhole-based confocal microscope applicable to femtosecond(fs)laser excitation.It can be employed not only for NIR-IIfluorescence intensity imaging,but also for multi-channelfluorescence lifetime imaging to recognize different structures with similarfluorescence spectrum.Moreover,it can be facilely combined with multiphotonfluorescence microscopy.A single fs pulsed laser is utilized to achieve up-conversion(visible multiphotonfluorescence)and down-conversion(NIR-II one-photonfluorescence)excitation simultaneously,extending imaging spectral channels,and thus facilitates multi-structure and multi-functional observation.

Relaxin-encapsulated polymeric metformin nanoparticles remodel tumor immune microenvironment by reducing CAFs for efficient triple-negative breast cancer immunotherapy

Cancer-associated fibroblasts(CAFs)are one of the most abundant stromal cells in the tumor microenvironment which mediate desmoplastic response and are the primary driver for an immunosuppressive microenvironment,leading to the failure of triple-negative breast cancer(TNBC)immunotherapy.Therefore,depleting CAFs may enhance the effect of immunotherapy(such as PD-L1 antibody).Relaxin(RLN)has been demonstrated to significantly improve transforming growth factor-β(TGF-β)induced CAFs activation and tumor immunosuppressive microenvironment.However,the short half-life and systemic vasodilation of RLN limit its in vivo efficacy.Here,plasmid encoding relaxin(pRLN)to locally express RLN was delivered with a new positively charged polymer named polymeric metformin(PolyMet),which could increase gene transfer efficiency significantly and have low toxicity that have been certified by our lab before.In order to improve the stability of pRLN in vivo,this complex was further formed lipid poly-γ-glutamic acid(PGA)/PolyMetpRLN nanoparticle(LPPR).The particle size of LPPR was 205.5±2.9 nm,and the zeta potential was+55.4±1.6 mV.LPPR displayed excellent tumor penetrating efficacy and weaken proliferation of CAFs in 4T1luc/CAFs tumor spheres in vitro.In vivo,it could reverse aberrantly activated CAFs by decreasing the expression of profibrogenic cytokine and remove the physical barrier to reshape the tumor stromal microenvironment,which enabled a 2.2-fold increase in cytotoxic T cell infiltration within the tumor and a decrease in immunosuppressive cells infiltration.Thus,LPPR was observed retarded tumor growth by itself in the 4T1 tumor bearing-mouse,and the reshaped immune microenvironment further led to facilitate antitumor effect when it combined with PD-L1 antibody(aPD-L1).Altogether,this study presented a novel therapeutic approach against tumor stroma using LPPR to achieve a combination regimen with immune checkpoint blockade therapy against the desmoplastic TNBC model.

A numerical study of mixing intensification for highly viscous fluids in multistage rotor-stator mixers

How to achieve uniform mixing of highly viscous fluids with low energy consumption is a major industry demand and one of the hot spots of mixing research.A typical multistage rotor-stator mixer(MRSM)equipped with a distributor was investigated to disclose the effects on the mixing performance and power consumption for highly viscous fluids via numerical simulation,considering the influence factors associated with different geometric parameters of both MRSM and the distributor.The mixing index and power consumption are used to evaluate the performance of the mixers.The dimensionless correlations for the mixing index and the power consumption are established considering the factors including the flow rate,rotor speed,the number of mixing units.Adopting the optimized mixer with the distributor(X1-T1),the mixing index increases to 0.85(obviously higher than 0.46 for the mixer T1 without a distributor),meanwhile the corresponding power consumption is about 1/5 of that of T1 achieving the same mixing effect.It illustrates that the distributor can significantly improve the mixing of highly viscous fluids in the MRSM without the cost of large power consumption.These results would provide a guidance on the design and optimization of multistage rotor-stator mixers in industrial applications.

GABAergic Interneuron Cell Therapy for Drug-Resistant Epilepsy

Epilepsy,one of the most common central nervous system disorders,affects about 70 million people of all ages worldwide[1].It is characterized by hyperactive neuronal networks,resulting from imbalanced excitatory and inhibitory activity.Despite the development of multiple next-generation anti-seizure medicines,one-third of epilepsy patients are still drug-resistant[2].

Adult-born neurons in critical period maintain hippocampal seizures via local aberrant excitatory circuits

Temporal lobe epilepsy(TLE),one common type of medically refractory epilepsy,is accompanied with altered adult-born dentate granule cells(abDGCs).However,the causal role of abDGCs in recurrent seizures of TLE is not fully understood.Here,taking advantage of optogenetic and chemogenetic tools to selectively manipulate abDGCs in a reversible manner.

Predicting the utilization factor of blasthole in rock roadways by random forest

The utilization factor of blasthole is a crucial indicator of the effectiveness of blasting in rock roadways.A significant value means that the explosive energy is fully utilized,the single-cycle advance is high,and the excavation rate is fast.A good blasting programme is a prerequisite for improving the utilization rate and predicting the utilization rate before blasting operations can verify the feasibility of the blasting programme.Firstly,a database of rock roadway blasting covering different geological and production conditions is estab-lished.Secondly,error analysis and the Gini coefficient method are used to weight the characteristic variables,quantify the importance of the variables and identify eight key indicators affecting the blasting hole utilization rate.Then,a random forest algorithm-based model for predicting utilization factor of blasthole is proposed,and the results of the model on the test set are:root mean square error(RMSE)is 0.0137,mean absolute error(MAE)is 0.0087,and coefficient of determination(R^(2))is 0.905.The performance of this method is com-pared with that of the neural network and support vector machine models on the test sets to verify the superiority of the random forest algorithm.Finally,to verify the generalization ability and practicality of the random forest prediction model,the model is applied to the rock roadway blasting construction of Gu Bei coal mine in Anhui Province.The results show that R2 is 0.913,so the model is reliable and accurate,which can meet the actual engineering requirements and lay the foundation for the promotion and application of this technology.

Helicobacter pylori infection may result in poor gastric cleanliness in magnetically controlled capsule gastroscopy examination: A singlecenter retrospective study

Objective:Magnetically controlled capsule gastroscopy(MCCG)is an effective method for screening gastric diseases;however,its performance may be affected by gastric cleanliness.We aimed to explore the correlation between Helicobacter pylori infection and the degree of gastric cleanliness in the MCCG.Methods:This retrospective study enrolled 297 participants from October 2020 to April 2024 at Sir Run Run Shaw Hospital,Zhejiang University School of Medicine.Participant characteristics,MCCG examination results,and(13)C-urea breath test(C13-UBT)results were collected.The gastric cleanliness in MCCG examinations was assessed using a gastric cleanliness score.Binary logistic regression was used to analyze the relationships among participant characteristics,H.pylori infection,and gastric cleanliness.Chi-square tests and Fisher's exact tests were used to analyze the relationships among gastric lesions,H.pylori infection,and gastric cleanliness.Results:Among the participants,24.2%had H.pylori infection,and 17.5%had poor gastric cleanliness.Hypertension(odds ratio[OR]:2.63;95%confidence interval[CI]:1.36e5.09;p?0.004)was associated with a greater likelihood of H.pylori infection.H.pylori infection(OR:3.76;95%CI:1.99e7.09;p<0.001)was an independent risk factor for poor gastric cleanliness in the MCCG.A significant disparity was noted in the prevalence of focal erosions(p<0.001),gastric ulcers(p?0.001),and positive gastric lesions(p?0.027)between the 2 groups with and without H.pylori infection.The proportion of positive gastric lesions was not significantly different between the good gastric cleanliness group and the poor gastric cleanliness group(25.7%vs.21.2%;p?0.490).Conclusion:The findings of this study revealed that H.pylori infection was associated with hypertension.H.pylori infection may lead to poor gastric cleanliness.Institutions are advised to perform C13-UBT before MCCG,and participants should be informed of the risk of poor gastric cleanliness if the results are positive.The decision to perform H.pylori eradication before MCCG should take into account patient willingness and the benefit-to-risk ratio.

矿物元素及炎症因子水平对慢性肾衰竭患者并发脑血管疾病的预测价值

目的 :明确矿物元素及炎症因子水平对慢性肾衰竭患者发生脑血管疾病事件的预测价值。方法 :连续性收集2016年1月~2020年6月间我院肾脏内科病房的慢性肾脏衰竭患者共389例,其中并发脑血管疾病患者236例作为研究组,未并发脑血管疾病患者153例作为对照组。采集两组患者静脉血作为检验样本,每个月检测血清中炎症因子和矿物元素指标水平并记录,为期12个月,最终各项数据取随访期间各项检查指标结果的平均值。将所检各指标纳入单因素分析和Logistic回归分析,最终筛选出影响慢性肾衰竭脑血管病发生的独立危险因素,并分析炎症因子和矿物元素与脑血管病发生的相关性。结果 :单因素分析结果显示,研究组动脉粥样硬化发生率高于对照组;IL-1、IL-6、TNF-α、Ca、P高于对照组;Mg、Se、Zn、Cu、Mn低于对照组。两组患者在年龄、性别构成、慢性肾衰竭病程以及原发病构成上无显著差异。多因素回归分析结果显示,IL-1、IL-6、TNF-α、Ca、P、Mg、Se、Zn、Cu、Mn均为慢性肾衰竭患者脑血管病发生的独立危险因素,且与其显著相关。结论 :慢性肾衰竭患者血液中相关炎症因子以及矿物元素,与脑血管疾病和动脉粥样硬化发生率呈显著相关,对慢性肾衰竭并发脑血管疾病有预测价值,并推测可用于慢性肾衰竭相关脑血管疾病的风险分层和作为潜在的治疗靶点。

Designing specific bacterial 16S primers to sequence and quantitate plant endo-bacteriome

Plant endophytic bacteria colonize the internal tissues of plants and interact with plants closely.The past two decades have witnessed the increasing application of next-generation 16S r RNA gene sequencing in the investigation of bacterial communities.However,deciphering plant endo-bacterial communities by this method is difficult because of the co-amplification of massive plant organellar DNAs with bacterial 16S.Here,we designed polymerase chain reaction(PCR) primer sets,including799F/1107R,322F/796R,and 322F-Dr/796Rs(primer pair 322F/796R with a penultimate-base substitution in 322F),that can specifically amplify bacterial 16S from plant total DNAs.We computationally and experimentally evaluated the specificity,coverage,and accuracy of the newly designed primer sets.Both 799F/1107R and 322F-Dr/796Rs produced plant DNA-free 16S amplicon libraries or reduced plant DNA contamination to lower than 5% for the plant materials with extremely-low-abundance bacterial communities.The primer set 322F-A/796R was used through absolute quantitative PCR to quantitate the population size of rice leaf or root endo-bacteriome,which revealed 10^(6)–10^(7) and 10^(9)–10^(10) bacteria per gram fresh weight,respectively.These 16S primer sets and amplification methods enable the simple and inexpensive next-generation sequencing and quantification of plant endo-bacteriome,which will significantly advance studies on the plant-related microbiome.

Impact of biochar amendment on soil aggregation varied with incubation duration and biochar pyrolysis temperature

Soil aggregation is one of the crucial processes that facilitate carbon sequestration and maintain soil fertility.So far,the effect of biochar amendment on soil aggregation remains inconclusive.Here,we tested the hypothesis that the response of soil aggregation to biochar addition varied with incubation duration and biochar chemistry.A one year microcosm experiment of soil with biochar was conducted that included biochar produced at three different temperatures(300,450,and 600°C),and three biochar application rates,i.e.,0,1,and 3 wt%.It was observed that after one and three months,biochar mainly(>90%)distributed in the micro-aggregates,and slightly reduced aggregate stability and increased proportion of micro-aggregates,which was demonstrated to result from the mechanical mixture of amended biochar with soil.Contrastingly,when the duration was prolonged to six months and one year,a significant increase in macro-aggregates(6.6-38.5%)and aggregate stability(7.3-29.4%)was detected,with the increasing extent being apparently higher for low-temperature biochar.This was related to the comparatively strong interaction of biochar particles with soil minerals or microbes after long-time incubation.The strong interaction was directly supported by the significant increase in H/C,O/C ratios of isolated biochar from treated soils,the detection of typical soil mineral elements on the surface of isolated biochar,and the increase in microbial biomass carbon of treated soils.The findings of this study highlighted the role of biochar type and amendment duration in mediating the effect of biochar application on soil aggregation.

Reactive glia-to-GABAergic neuron reprogramming:a“golden touch”strategy to alleviate intractable seizures

This commentary highlights a research article published recently in Cell Stem Cell“Reprogramming reactive glia into interneurons reduces chronic seizure activity in a mouse model of mesial temporal lobe epilepsy”.Generally,Lentini et al.reveal a strategy to fulfill in vivo glia-to-neuron reprogramming,which is a potential disease-modifying strategy for treatment of intractable seizures.Here,we describe exciting research advances in the treatment of intractable seizures based on this research article,summarizing its key findings,emphasizing its importance and providing further discussions.Further,issues worthy of further investigations are also postulated so that clinic translation can be better achieved.