Breast cancer is reported to be one of the most lethal cancers in women, and its multi-target detection can help improve the accuracy of diagnosis. In this work, a cluster regularly interspaced short palindromic repeats (CRISPR)-Cas13a/Cas12a-based system was established for the simultaneous fluorescence detection of breast cancer biomarkers circROBO1 and BRCA1. CRISPR-Cas13a and CRISPR-Cas12a were directly activated by their respective targets, resulting in the cleavage of short RNA and DNA reporters, respectively, thus the signals of 6-carboxyfluorescein (FAM) and 6-carboxy-xrhodamine (ROX) were restored. As the fluorescence intensities of FAM and ROX were dependent on the concentrations of circROBO1 and BRCA1, respectively, synchronous fluorescence scanning could achieve one-step detection of circROBO1 and BRCA1 with detection limits of 0.013 pM and 0.26 pM, respectively. The system was highly sensitive and specific, holding high diagnostic potential for the detection of clinical samples. Furthermore, the competing endogenous RNA mechanism between circROBO1 and BRCA1 was also explored, providing a reliable basis for the intrinsic regulatory mechanism of breast cancer.

Stimuli-responsive behaviors and controlled release in liposomes are pivotal in nanomedicine. To this end, we present an approach using a photoresponsive azobenzene nanocluster (AzDmpNC), prepared from azobenzene compounds through melting and aggregation. When integrated with liposomes, they form photoresponsive vesicles. The morphology and association with liposomes were investigated by using transmission electron microscopy. Liposomes loaded with calcein exhibited a 9.58% increased release after UV exposure. To gain insights into the underlying processes and elucidate the mechanisms involved. The molecular dynamic simulations based on the reactive force field and all-atom force field were employed to analyze the aggregation of isomers into nanoclusters and their impacts on phospholipid membranes, respectively. The results indicate that the nanoclusters primarily aggregate through π-π and T-stacking forces. The force density inside the cis-isomer of AzDmpNC formed after photoisomerization is lower, leading to its easier dispersion, rapid diffusion, and penetration into the membrane, disrupting the densification.

Successful construction of a detection method for Salmonella typhimurium (S. typhimurium) based on the synergy of hybridization chain reaction (HCR) and fluorescence was realized in this paper. First, the aptamer modified with the quenching group Black Hole Quencher-1 acid (BHQ1) was immobilized on the magnetic beads in combination with the complementary chain of the aptamer modified with 6-carboxyfluorescein (6-FAM). Second, S. typhimurium and cDNA-6-FAM immobilized on magnetic beads competitively bound to the aptamer. Finally, the cDNA-6-FAM was released after magnetic separation acted as a promoter to trigger HCR amplification when the target presented. The fluorescence signal could be significantly improved by the combination of green SYBR Green I (SGI) and HCR long double-stranded DNA and the fluorescent synergy of 6-FAM and SGI. Because of the separation of target and its aptamer, the trigger strand was abstracted by magnetic separation. There was no HCR to generate long double-stranded DNA, and the fluorescence of excess hairpin/SGI could be adsorbed through UIO66 so that only a very low background signal was detected. This fluorescent sensor was capable of monitoring S. typhimurium in the range of 10-3.2 × 107 CFU mL-1 with a limit of detection as low as 1.5 CFU mL-1. Because of the excellent properties of the aptasensor and the validity of SGI fluorescence synergy, this HCR enzyme-free amplification strategy could be generalized to other areas.

BACKGROUND: Human amniotic mesenchymal stem cells (hAMSCs) derived from amniotic membrane have multilineage differentiation, immunosuppressive, and anti-inflammation which makes them suitable for the treatment of various diseases.
OBJECTIVE: This study aimed to explore the therapeutic effect and molecular mechanism of hAMSCs in ventricular remodeling (VR).
METHODS: hAMSCs were characterized by a series of experiments such as flow cytometric analysis, immunofluorescence, differentiative induction and tumorigenicity. Mouse VR model was induced by isoproterenol (ISO) peritoneally, and the therapeutic effects and the potential mechanisms of hAMSCs transplantation were evaluated by echocardiography, carboxy fluorescein diacetate succinimidyl ester (CFSE) labeled cell tracing, histochemistry, qRT-PCR and western blot analysis. The co-culturing experiments were carried out for further exploring the mechanisms of hAMSCs-derived conditioned medium (CM) on macrophage polarization and fibroblast fibrosis in vitro.
RESULTS: hAMSCs transplantation significantly alleviated ISO-induced VR including cardiac hypertrophy and fibrosis with the improvements of cardiac functions. CFSE labeled hAMSCs kept an undifferentiated state in heart, indicating that hAMSCs-mediated the improvement of ISO-induced VR might be related to their paracrine effects. hAMSCs markedly inhibited ISO-induced inflammation and fibrosis, seen as the increase of M2 macrophage infiltration and the expressions of CD206 and IL-10, and the decreases of CD86, iNOS, COL3 and αSMA expressions in heart, suggesting that hAMSCs transplantation promoted the polarization of M2 macrophages and inhibited the polarization of M1 macrophages. Mechanically, hAMSCs-derived CM significantly increased the expressions of CD206, IL-10, Arg-1 and reduced the expressions of iNOS and IL-6 in RAW264.7 macrophages in vitro. Interestingly, RAW264.7-CM remarkably promoted the expressions of anti-inflammatory factors such as IL-10, IDO, and COX2 in hAMSCs. Furthermore, the CM derived from hAMSCs pretreated with RAW264.7-CM markedly inhibited the expressions of fibrogenesis genes such as αSMA and COL3 in 3T3 cells.
CONCLUSIONS: Our results demonstrated that hAMSCs effectively alleviated ISO-induced cardiac hypertrophy and fibrosis, and improved the cardiac functions in mice, and the underlying mechanisms might be related to inhibiting the inflammation and fibrosis during the ventricular remodeling through promoting the polarization of CD206hiIL-10hi macrophages in heart tissues. Our study strongly suggested that by taking the advantages of the potent immunosuppressive and anti-inflammatory effects, hAMSCs may provide an alternative therapeutic approach for prevention and treatment of VR clinically.

This study investigated the relationship between bacterial growth, viability, and extracellular polymeric substances (EPS) formation in biofilms, particularly regarding resistance development. It also examined the impact of chemical factors on the EPS matrix and bacterial proliferation in oral biofilms.
Three multi-species oral biofilms were incubated in anaerobic conditions. Three strains of Enterococcus faecalis were incubated in aerobic conditions. The incubation periods ranged from 0 h to 7 days for short-term biofilms, and from 3 to 90 days for long-term biofilms. Fluorescent labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) and flow cytometry were used to track EPS and bacterial growth. Confocal laser scanning microscopy (CLSM) assessed bacterial viability and EPS structure. Biofilms aged 7, 14, and 21 days were treated with 2 % chlorhexidine (CHX) and 1 % sodium hypochlorite (NaOCl) to evaluate their effects on EPS and bacterial proliferation.
Short-term biofilms showed rapid bacterial proliferation and a gradual increase in EPS, maintaining stable viability. In the first two weeks, a significant rise in CFSE indicated growing maturity. From 14 to 90 days, EPS and CFSE levels stabilized. Following treatment, CHX significantly reduced bacterial proliferation, while NaOCl decreased EPS volume.
Biofilm development involves a balance between bacterial proliferation and EPS production. The complexity of this process poses challenges in treating biofilm-associated infections, requiring strategies tailored to the biofilm\'s developmental stage.
For effective root canal treatment, it is imperative to focus on reducing bacterial proliferation during the early stages of oral infections. In contrast, strategies aimed at minimizing EPS production could be more beneficial for long-term management of these conditions.

Objective: To evaluate the efficacy of 0.05% cyclosporine A eye drops combined with vitamin A palmitate eye gel in the treatment of dry eye associated with meibomian gland dysfunction (MGD). Methods: A single-center, prospective, randomized, parallel controlled trial design was used to include patients diagnosed with MGD-associated dry eye. The patients were randomly divided into three groups and administered with medications binocularly for 12 weeks. The CsA+VA group was given 0.05% cyclosporine A eye drops twice a day and vitamin A palmitate eye gel three times a day. The CsA+HA group was given 0.05% cyclosporine A eye drops twice a day and 0.1% sodium hyaluronate eye drops three times a day. The HA group was given 0.1% sodium hyaluronate eye drops 3 times a day. The OSDI score, tear meniscus height, fluorescein tear break-up time, Schirmer Ⅰ test (without anesthesia), tear film lipid layer thickness, meibomian gland morphology and function examination, and corneal fluorescein sodium staining score were evaluated at baseline, 4, 8, and 12 weeks after the initiation of the treatment, respectively. Results: A total of 120 patients with MGD-related dry eye met the enrollment criteria, but 10 patients were lost to follow-up; 110 patients were finally included for observation, including 36 patients in the CsA+VA group, 38 in the CsA+HA group and 36 in the HA group. The OSDI score, tear meniscus height, fluorescein tear break-up time and meibomian gland secretion of the 3 groups were significantly improved. At the 12th week of the treatment, the differences of the CsA+VA group [25.45±15.11, (0.30±0.13) mm, (3.72±1.40) s, (5.03±2.52) points] and the CsA+HA group [26.98±16.89, (0.27±0.10) mm, (4.34±1.76) s, (5.11±2.39) points] from the HA group [24.57±11.26, (0.24±0.06) mm, (3.18±1.11) s, (9.11±3.34) points] were statistically significant (P<0.05). Compared with the CsA+HA group [(68.39±26.66) nm], the tear film lipid layer thickness in the CsA+VA group [(72.61±23.65) nm] was significantly increased (P<0.05). In the CsA+VA group, the meibomian gland secretion characters and discharge capacity among patients with severe abnormalities [(6.28±2.59) and (5.89±2.77) points at the 12th week of treatment], moderate abnormalities [(4.27±2.02) and (4.64±2.02) points at the 12th week of treatment] and mild abnormalities [(2.80±0.84) and (2.60±0.55) points at the 12th week of treatment] were significantly different (P<0.05). Conclusion: 0.05% cyclosporine A combined with vitamin A palmitate can significantly improve the symptoms and signs of patients with MGD-related dry eye, especially the tear film lipid layer thickness and the meibomian gland secretion characters and discharge capacity in severe cases.
目的： 评价0.05%环孢素A联合维生素A棕榈酸酯治疗睑板腺功能障碍（MGD）相关干眼的疗效。 方法： 单中心、前瞻性、随机平行对照试验。纳入2022年7至11月在首都医科大学附属北京同仁医院北京同仁眼科中心确诊为MGD相关干眼患者，采用随机数字表法随机分为CsA+VA组（0.05%环孢素A滴眼液双眼点药，每日2次；维生素A棕榈酸酯眼用凝胶双眼点药，每日3次）、CsA+HA组（0.05%环孢素A滴眼液双眼点药，每日2次；0.1%玻璃酸钠滴眼液双眼点药，每日3次）、HA组（0.1%玻璃酸钠滴眼液双眼点药，每日3次）3个组治疗12周，分别记录并比较治疗前（基线）以及治疗后第4、8、12周的眼表疾病指数（OSDI）、泪河高度、荧光素泪膜破裂时间、Schirmer Ⅰ试验结果（无麻醉）、泪膜脂质层厚度、睑板腺形态、睑板腺分泌物排出能力和性状评分、角膜荧光素染色评分。采用单因素方差分析、单因素重复测量方差分析、两因素重复测量方差分析、Bonferroni检验等进行统计学分析。 结果： 共收集120例（120只眼）符合标准的MGD相关干眼患者，失访10例（10只眼），最终纳入研究患者110例（110只眼），年龄为22~73岁，男性22例（22只眼），女性88例（88只眼）；其中CsA+VA组38例（38只眼），CsA+HA组36例（36只眼），HA组36例（36只眼）。3个组OSDI、泪河高度、荧光素泪膜破裂时间和睑板腺分泌物性状评分治疗后均有明显改善，其中治疗第12周，CsA+VA组［25.45±15.11，（0.30±0.13）mm，（3.72±1.40）s，（5.03±2.52）分］和CsA+HA组［26.98±16.89，（0.27±0.10）mm，（4.34±1.76）s，（5.11±2.39）分］与HA组［24.57±11.26，（0.24±0.06）mm，（3.18±1.11）s，（9.11±3.34）分］相比，差异均有统计学意义（均P<0.05）；CsA+VA组的泪膜脂质层厚度［（72.61±23.65）nm］明显高于CsA+HA组［（68.39±26.66）nm］（P<0.05）。在CsA+VA组中，睑板腺分泌物性状和排出能力重度异常者［治疗第12周，（6.28±2.59）和（5.89±2.77）分］、中度异常者［治疗第12周，（4.27±2.02）和（4.64±2.02）分］和轻度异常者［治疗第12周，（2.80±0.84）和（2.60±0.55）分］的睑板腺分泌物性状评分和排出能力评分比较，差异均有统计学意义（均P<0.05）。 结论： 0.05%环孢素A联合维生素A棕榈酸酯可明显改善MGD相关干眼患者的症状和体征，尤其可明显改善泪膜脂质层厚度和重度患者的睑板腺分泌物性状和排出能力。.

Unilateral high myopia (uHM), commonly observed in patients with retinal diseases or only with high myopia, is frequently associated with amblyopia with poor prognosis. This study aims to reveal the clinical and genetic spectrum of uHM in a large Chinese cohort.
A total of 75 probands with simplex uHM were included in our Pediatric and Genetic Eye Clinic. Patients with significant posterior anomalies other than myopic fundus changes were excluded. Variants were detected by exome sequencing and then analyzed through multiple-step bioinformatic and co-segregation analysis and finally confirmed by Sanger sequencing. Genetic findings were correlated with associated clinical data for analysis.
Among the 75 probands with a mean age of 6.21 ± 4.70 years at the presentation, myopic fundus of C1 and C2 was observed in 73 (97.3%) probands. Surprisingly, specific peripheral changes were identified in 63 eyes involving 36 (48.0%) probands after extensive examination, including peripheral retinal avascular zone (74.6%, 47/63 eyes), neovascularization (54.0%), fluorescein leakage (31.7%), peripheral pigmentary changes (31.7%), and others. Exome sequencing identified 21 potential pathogenic variants of 13 genes in 20 of 75 (26.7%) probands, including genes for Stickler syndrome (COL11A1 and COL2A1; 6/20), FEVR (FZD4, LRP5, and TSPAN12; 5/20), and others (FBN1, GPR179, ZEB2, PAX6, GPR143, OPN1LW, FRMD7, and CACNA1F; 9/20). For the peripheral retinal changes in the 20 probands, variants in Stickler syndrome-related genes were predominantly associated with retinal pigmentary changes, lattice degeneration, and retinal avascular region, while variants in genes related to FEVR were mainly associated with the avascular zone, neovascularization, and fluorescein leakage.
Genetic defects were identified in about one-fourth of simplex uHM patients in which significant consequences may be hidden under a classic myopic fundus in up to half. To our knowledge, this is the first systematic genetic study on simplex uHM to date. In addition to routine care of strabismus and amblyopia, careful examination of the peripheral retina and genetic screening is warranted for patients with uHM in order to identify signs of risk for retinal detachment and other complications and provide meaningful genetic counseling.

In this paper, we synthesized silver nanoclusters using bovine serum albumin (BSA) as a template (BSA@AgNCs). Then, we anchored hydroxyphenyl fluorescein (HPF) to yield HPF-BSA@AgNCs. When exposed to X-rays, hydroxyl (∙OH) radicals generated by radiolysis of water react with HPF to produce fluorescein, which emits enhanced fluorescence at 515 nm (λex = 480 nm). The fluorescence intensity of BSA@AgNCs at 685 nm (λex = 480 nm) remains stable when exposed to X-rays. This HPF-BSA@AgNCs ratiometric fluorescence sensor can rapidly detect 0.1-20 Gy (the energy deposited per unit mass, J/kg) of X-rays. In addition, HPF-BSA@AgNCs exhibit good durability and temperature stability. Finally, HPF-BSA@AgNCs were used to measure the absorbed doses of A549 cells and evaluate the cell irradiation damage.

Liposomes have emerged as versatile nanocarriers, finding applications not only in drug delivery but also in pathogen detection and diagnostics. This study aimed to enhance the sensitivity of liposomes to Staphylococcus aureus by investigating the impact of lipid composition on liposomes loaded with 5(6)-carboxyfluorescein (CF). Liposomes were fabricated using various concentrations of cholesterol (10-40 mol%) combined with saturated phospholipids. Dynamic light scattering results revealed that higher cholesterol concentrations led to reduced liposome size, CF release (%), and entrapment efficiency (%). Liposome sensitivity towards S. aureus was evaluated by using CF-loaded liposomes with and without aptamer insertion. Liposomes with a higher cholesterol content (40 mol%) exhibited a strong ability to detect low bacterial concentrations down to 5 × 102 CFU/mL without relying solely on specific receptor-ligand recognition. However, functionalizing the liposome with an aptamer further improved the specificity and sensitivity of S. aureus detection at even lower concentrations, down to 80 CFU/mL, in the wide range of 80-107 CFU/mL. This study highlights the potential for optimizing the lipid composition of liposomes to improve their sensitivity for pathogen detection, particularly when combined with aptamer-based strategies.

There is an urgent need to assess material degradation in situ and in real time for their promising application in regeneration therapy. However, traditional monitoring methods in vitro cannot always profile the complicated behavior in vivo. This study designed and synthesized a new biodegradable polyurethane (PU-P) scaffold with polycaprolactone glycol, isophorone diisocyanate, and l-lysine ethyl ester dihydrochloride. To monitor the degradation process of PU-P, calcein was introduced into the backbone (PU-5) as a chromophore tracing in different sites of the body and undegradable fluorescent scaffold (CPU-5) as the control group. Both PU-P and PU-5 can be enzymatically degraded, and the degradation products are molecularly small and biosafe. Meanwhile, by virtue of calcein anchoring with urethane, polymer chains of PU-5 have maintained the conformational stability and extended the system conjugation, raising a structure-induced emission effect that successfully achieved a significant enhancement in the fluorescence intensity better than pristine calcein. Evidently, unlike the weak fluorescent response of CPU-5, PU-5 and its degradation can be clearly imaged and monitored in real time after implantation in the subcutaneous tissue of nude mice. Meanwhile, the in situ osteogeneration has also been promoted after the two degradable scaffolds have been implanted in the rabbit femoral condyles and degraded with time. To sum up, the strategy of underpinning tracers into degradable polymer chains provides a possible and effective way for real-time monitoring of the degradation process of implants in vivo.