Single-cell analysis of phenotypic plasticity could improve the development of more effective therapeutics. Still, the development of tools to measure single-cell heterogeneity has lagged due to difficulties in manipulating and culturing single cells. Here, we describe a single-cell culture and phenotyping platform that employs a starburst microfluidic network and automatic liquid handling system to capture single cells for long-term culture and multi-dimensional analysis and quantify their clonal properties via their surface biomarker and secreted cytokine/growth factor profiles. Studies performed on this platform found that cells derived from single-cell cultures maintained phenotypic equilibria similar to their parental populations. Single-cell cultures exposed to chemotherapeutic drugs stochastically disrupted this balance to favor stem-like cells. They had enhanced expression of mRNAs and secreted factors associated with cell signaling, survival, and differentiation. This single-cell analysis approach can be extended to analyze more complex phenotypes and screen responses to therapeutic targets.

Objective To investigate the distribution of dense fine speckled(DFS)antinuclear antibody(ANA)in different ages,genders,and diseases,and to explore its clinical significance in the diagnosis of auto-immune disease(AID).Methods A retrospective summary was conducted on the clinical basic data of 53520 patients who visited the hospital from January 2022 to April 2024,as well as the results of indirect immunoflu-orescence(IIF)ANA fluorescence karyotype and titer,and immunoblotting ANA spectrum,and the relation-ship between DFS positivity and AID was analyzed.Results Of the 53 520 serum samples,the rate of ANA fluorescence positivity was 32.40％,with DFS-positive samples accounting for 1.18％of all ANA fluores-cence-positive samples.The male-to-female ratio of DFS positives was 1∶1.94,with the highest percentage of 51.9％in the 21-40 age group.Among DFS positive individuals,only 12.14％were clinically diagnosed with AID disease,significantly lower than other ANA fluorescent karyotype positive individuals(23.31％,P＜0.001).The overall positivity rate of ANA spectra using immunoblotting in DFS-positive individuals was 23.54％,which was significantly lower than that in other fluorescent karyotype positive ANA individuals(73.26％,P＜0.001).Moreover,DFS positive individuals were mainly accompanied by antibody positivity such as anti-Ro52 antibodies,which had no diagnostic specificity in AID diagnosis.When comparing DFS posi-tive individuals with similar fluorescence morphology to those with homogeneous nuclear granules and nuclei,at an ANA titer of 1∶100,the proportion of AID patients in homogeneous nuclear granules and nuclei was 20.04％,significantly higher than the proportion of AID patients in DFS positive individuals(9.44％,P=0.001).With the increase of ANA titer,the proportion of AID diseases significantly increased in both karyo-type samples.When the ANA titer of DFS positive samples was increased to ≥ 1∶320,the proportion of AID diseases significantly increased to 30.77％(P=0.002).Conclusion DFS is a rare ANA fluorescent karyo-type,and DFS positive individuals are mainly found in females aged 21-40,mostly non AID patients.Howev-er,high titers of DFS cannot rule out the possibility of AID.In clinical work,special attention should be paid to distinguishing it from the homogeneous nuclear granule type to avoid misdiagnosis.

Objective To identify the pathogen causing bloodstream infection(BSI)in an elderly patient and to de-scribe its pathogenic characteristics.This study also reviews recent relevant literatures on Trichoderma longibra-chiatum(T.longibrachiatum)infections.Methods Blood specimens from central and peripheral veins were co-llected for blood culture.The morphological characteristics and drug susceptibility of the isolated filamentous fungus were analyzed.The sequences of the isolated strains were detected using the internal transcribed spacer(ITS)re-gion,and then analyzed using MEGA software.Results Both morphological observation and ITS sequencing su-ggested that the pathogen was T.longibrachiatum.The minimum inhibitory concentration(MIC)against the strain detected in accordance with the M38-A2 Guideline for the Drug Susceptibility of Filamentous Fungi issued by the Clinical and Laboratory Standards Institute(CLSI)of the United States showed that the MICs of micafungin,caspofungin,5-fluorocytosine,posaconazole,voriconazole,itraconazole,fluconazole,anidulafungin and amphoteri-cin were 8,8,64,2,0.5,2,64,8 and 4 μg/mL respectively.The patient recovered well after anti-fungal treat-ment with voriconazole.A total of 15 cases of T.longibrachiatum infection were reported in the literature.The main underlying disease was transplantation(n=5),and the main clinical manifestation was pulmonary infection(n=5).Of these,9 cases survived,3 died,and 3 had an unknown prognosis.Conclusion T.longibrachiatum in-fection is more common in patients with low immunity and more underlying diseases,with a high fatality rate,lack standardized clinical treatment guidelines,lack break points in drug sensitivity results,and is resistant to common antifungal drugs.It is of guiding significance to isolate and identify pathogens in time and provide corresponding drug sensitivity testing results for clinical drug use.

Objective To identify the pathogen causing bloodstream infection(BSI)in an elderly patient and to de-scribe its pathogenic characteristics.This study also reviews recent relevant literatures on Trichoderma longibra-chiatum(T.longibrachiatum)infections.Methods Blood specimens from central and peripheral veins were co-llected for blood culture.The morphological characteristics and drug susceptibility of the isolated filamentous fungus were analyzed.The sequences of the isolated strains were detected using the internal transcribed spacer(ITS)re-gion,and then analyzed using MEGA software.Results Both morphological observation and ITS sequencing su-ggested that the pathogen was T.longibrachiatum.The minimum inhibitory concentration(MIC)against the strain detected in accordance with the M38-A2 Guideline for the Drug Susceptibility of Filamentous Fungi issued by the Clinical and Laboratory Standards Institute(CLSI)of the United States showed that the MICs of micafungin,caspofungin,5-fluorocytosine,posaconazole,voriconazole,itraconazole,fluconazole,anidulafungin and amphoteri-cin were 8,8,64,2,0.5,2,64,8 and 4 μg/mL respectively.The patient recovered well after anti-fungal treat-ment with voriconazole.A total of 15 cases of T.longibrachiatum infection were reported in the literature.The main underlying disease was transplantation(n=5),and the main clinical manifestation was pulmonary infection(n=5).Of these,9 cases survived,3 died,and 3 had an unknown prognosis.Conclusion T.longibrachiatum in-fection is more common in patients with low immunity and more underlying diseases,with a high fatality rate,lack standardized clinical treatment guidelines,lack break points in drug sensitivity results,and is resistant to common antifungal drugs.It is of guiding significance to isolate and identify pathogens in time and provide corresponding drug sensitivity testing results for clinical drug use.

Nuclear medicine plays an indispensable role in the diagnosis,treatment,and prognosis of a wide range of diseases.Nuclear medicine using radionuclides for diagnosis has the advantages of accuracy,speed,high sensitivity and high resolution.Currently,several radionuclides play pivotal roles in disease diagnosis.This article primarily examines the clinical application and research of diagnostic radionuclides,including 18 F,89 Zr,68 Ga,99m Tc,131 I,123 I,and 11 C.The objective is to offer valuable insights for disease diagnosis and staging of diseases.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

Objective:To explore the clinical feasibility of finger-pressing therapy based on the theory of treating impotence alone with Yang Ming to reduce incidence of ICU acquired weakness (ICU-AW) in critically ill children and provide a feasible nursing plan for ICU acquired asthenia in critically ill children.Methods:A quasi-experimental study was conducted. A total of 73 critically ill children were admitted to the PICU of Kunming Children′s Hospital from January 1 to April 30, 2021. According to the random number table, the subjects were divided into the observation group (37 cases) and the control group (36 cases). Children in the control group received routine PICU nursing. The children in the observation group were treated with PICU routine nursing and finger-pressing therapy based on the theory of treating impotence alone with Yang Ming. The two groups were compared in terms of limb muscle strength score (MRC-Score), incidence of ICU-AW, basic activities of life (Barthel Index, BI), limb muscle thickness.Results:After intervention, the MRC-Score of the observation group was 50 (46, 52) points, which was higher than 46 (40, 48) points of the control group, and the difference between the two groups was statistically significant ( Z=-3.70, P<0.05). The incidence of ICU-AW in the observation group was 32.43% (12/37), and the incidence of ICU-AW in the control group was 72.22% (26/36). The difference between the two groups was statistically significant ( χ2=11.58, P<0.05). The BI score of the observation group was 63 (50, 70), which was higher than 44 (40,60) of the control group, and the difference between the two groups was statistically significant ( Z=-3.94, P<0.05). The reduction degree of quadriceps femoris thickness in the observation group at D3-D1 was (-0.381 ± 0.131) cm, which was lower than (-0.762 ± 0.182) cm in the control group, and the difference between the two groups was statistically significant ( t=10.29, P<0.05). Conclusions:The application of finger-pressing therapy guided by theory of treating impotence alone with Yang Ming in the early rehabilitation of critically ill children can enhance muscle strength, prevent muscle atrophy and reduce the incidence of ICU-AW in critically ill children.

DNA is a biological polymer that encodes and stores genetic information in all living organism. Particularly, the precise nucleobase pairing inside DNA is exploited for the self-assembling of nanostructures with defined size, shape and functionality. These DNA nanostructures are known as framework nucleic acids (FNAs) for their skeleton-like features. Recently, FNAs have been explored in various fields ranging from physics, chemistry to biology. In this review, we mainly focus on the recent progress of FNAs in a pharmaceutical perspective. We summarize the advantages and applications of FNAs for drug discovery, drug delivery and drug analysis. We further discuss the drawbacks of FNAs and provide an outlook on the pharmaceutical research direction of FNAs in the future.

Stimulator of interferon genes (STING) is a cytosolic DNA sensor which is regarded as a potential target for antitumor immunotherapy. However, clinical trials of STING agonists display limited anti-tumor effects and dose-dependent side-effects like inflammatory damage and cell toxicity. Here, we showed that tetrahedral DNA nanostructures (TDNs) actively enter macrophages to promote STING activation and M1 polarization in a size-dependent manner, and synergized with Mn²⁺ to enhance the expressions of IFN-β and iNOS, as well as the co-stimulatory molecules for antigen presentation. Moreover, to reduce the cytotoxicity of Mn²⁺, we constructed a TDN-MnO₂ complex and found that it displayed a much higher efficacy than TDN plus Mn²⁺ to initiate macrophage activation and anti-tumor response both in vitro and in vivo. Together, our studies explored a novel immune activation effect of TDN in cancer therapy and its synergistic therapeutic outcomes with MnO₂. These findings provide new therapeutic opportunities for cancer therapy.

Objective:To explore the clinical effects of high flow nasal cannula (HighFlow) on patients with traumatic flail chest after removing mechanical ventilation.Methods:A retrospective analysis was carried out on 80 traumatic flail chest patients with mechanical ventilation, admitted to the ICU of Department of Emergency and Critical Care Medicine of Shanghai Changzheng Hospital from October 2016 to December 2019, who were divided into the HighFlow group and the conventional oxygen therapy (COT) group. The clinical effects of high flow nasal cannula on PaO 2/FiO 2 ratio, ventilation-free days, re-intubation case number, length of stay in ICU, incidence of ventilation associated pneumonia (VAP), and ICU mortality were analyzed. Results:The time point of extubation of the HighFlow group was (4.6±2.1) d, while that of the COT group was (4.3±2.2) d ( P>0.05). At the time point of extubation, the differences of the ventilation oxygenation indexes, i. e. arterial blood gas pH, PaO 2, and PaCO 2, heart rate (HR), and mean arterial pressure (MAP), between the two groups were not statistically significant ( P>0.05). PaO 2/FiO 2 and PaCO 2 at 2 h after extubation in the HighFlow group were significantly improved as compared with in the COT group; the re-intubation rate of the HighFlow group was significantly lower than that of the COT group; the time of ventilation-free of the HighFlow group was significantly longer than that of the COT group; the length of stay in ICU of the HighFlow group was significantly shorter than that of the COT group; the incidence of VAP of the HighFlow group was obviously lower than that of the COT group; all differences were statistically significant ( P<0.05). Conclusion:For traumatic flail chest patients with mechanical ventilation, high flow oxygen therapy can help them wean from mechanical ventilation earlier and reduce the incidence of VAP, which is worthy of popularization and application in clinic.