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The fluorescence high-throughput screening method is of importance for new antioxidant drug candidate discovery for the treatment of serious hepatorenal syndrome, which displayed an obvious upregulated peroxynitrite level. However, most of the current ONOO - probes possessed incomplete fluorescence quenching efficiency, which can result in non-negligible probe inherent fluorescence. Hence, we utilized the probe conjugated structure disruption strategy to construct hydrogenation phosphorus-substituted rhodamine ( H-PRh ) with "zero" probe inherent fluorescence character. Based on the precursor, a series of natural products were screened for identifying antioxidant drug candidates. Luteolin was screened out by activating the Sirt1-Nrf2-HO-1 signaling pathway to regulate the accumulation of ONOO - in the hepatorenal syndrome. Overall, the "zero" probe inherent fluorescence ONOO - sensor constructed here applies for a promising and versatile toolbox for illuminating the ONOO - -related pathological process in the hepatorenal syndrome. Besides, this strategy of constructing highly sensitive sensors could serve as a valuable reference for further fluorescent probes.

High-performance fluorescent probes stand as indispensable tools in fluorescence-guided imaging, and are crucial for precise delineation of focal tissue while minimizing unnecessary removal of healthy tissue. Herein, machine-learning-assisted strategy to investigate the current available xanthene dyes is first proposed, and a quantitative prediction model to guide the rational synthesis of novel fluorescent molecules with the desired pH responsivity is constructed. Two novel Si─rhodamine derivatives are successfully achieved and the cathepsin/pH sequentially activated probe Si─rhodamine─cathepsin-pH (SiR─CTS-pH) is constructed. The results reveal that SiR─CTS-pH exhibits higher signal-to-noise ratio of fluorescence imaging, compared to single pH or cathepsin-activated probe. Moreover, SiR─CTS-pH shows strong differentiation abilities for tumor cells and tissues and accurately discriminates the complex hepatocellular carcinoma tissues from normal ones, indicating its significant application potential in clinical practice. Therefore, the continuous development of xanthene dyes and the rational design of superior fluorescent molecules through machine-learning-assisted model broaden the path and provide more advanced methods to researchers., (© 2024 Wiley‐VCH GmbH.)

Activatable probes with a higher signal-to-background ratio and accuracy are essential for monitoring liver cancer as well as intraoperative fluorescence navigation. However, the presence of only one biomarker is usually not sufficient to meet the high requirement of a signal-to-background ratio in cancer surveillance, leading to the risk of misdiagnosis. In this work, a dual-locked activation response probe, Si-NTR-LAP , for nitroreductase and leucine aminopeptidase was reported. This dual-locked probe provides better tumor recognition and a higher signal-to-noise ratio than that of single-locked probes ( Si-LAP and Si-NTR ). In both the subcutaneous tumor model and the more complex orthotopic hepatocellular carcinoma model, the probe was able to identify tumor tissue with high specificity and accurately differentiate the boundaries between tumor tissue and normal tissue. Therefore, the dual-locked probe may provide a new and practical strategy for applying to real patient tumor tissue samples.

The separate vertical wire (SVW) technique and the improved candy box (CB) technique have been proposed for treating inferior pole patellar fractures. However, there is still a lack of clear explanation regarding the location of the wire passing through the patella. Five models of SVW techniques were established in different positions. Finite element analysis was then conducted to determine the optimal bone tunnel position for the SVW technique. Based on these findings, six groups of finite element models were created for CB techniques. The maximum displacement and stress on both the patella and steel wire were compared among these groups under 100-N, 200-N, 300-N, 400-N, and 500-N force loads. The results indicated that, in the SVW technique, the steel wire group near the fracture end of the longitudinal bone tunnel showed minimal displacement and stress on the patella when subjected to different forces. On the other hand, in the CB technique, both the patella and wire experienced minimal stress when a transverse bone tunnel wire was placed near the upper posterior aspect of patella. In conclusion, the SVW technique may require the bone tunnel wire to be positioned near the fractured end of the lower pole of the patella. On the other hand, in CB technique, the transverse bone tunnel wire passing through the patella may be close to its upper posterior aspect. However, further validation is necessary through comprehensive finite element analysis and additional biomechanical experiments., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Fan, Dang, Tan, Liu, Yang and Xiang.)

Enriching the palette of high-performance fluorescent dyes is vital to support the frontier of biomedical imaging. Although various rhodamine skeletons remain the premier type of small-molecule fluorophores due to the apparent high brightness and flexible modifiability, they still suffer from the inherent defect of small Stokes shift due to the nonideal fluorescence imaging signal-to-background ratio. Especially, the rising class of fluorescent dyes, sulfone-substituted xanthone, exhibits great potential, but low chemical stability is also pointed out as the problem. Molecular engineering of sulfone-xanthone to obtain a large Stokes shift and high stability is highly desired, but it is still scarce. Herein, we present the combination modification method for optimizing the performance of sulfone-xanthone. These redesigned fluorescent skeletons owned greatly improved stability and Stokes shift compared with the parent sulfone-rhodamine. To the proof of bioimaging capacity, annexin protein-targeted peptide LS301 was introduced to the most promising dyes, J-S-ARh, to form the tumor-targeted fluorescent probe, J-S-LS301. The resulting probe, J-S-LS301, can be an outstanding fluorescence tool for the orthotopic transplantation tumor model of hepatocellular carcinoma imaging and on-site pathological analysis. In summary, the combination method could serve as a basis for rational optimization of sulfone-xanthone. Overall, the chemistry reported here broadens the scope of accessible sulfone-xanthone functionality and, in turn, enables to facilitate the translation of biomedical research toward the clinical domain., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

The abnormal microenvironment parameter, viscosity, is closely connected with various diffusion processes, signal transduction, molecule interactions, and various diseases. It is greatly significant to design viscosity-dependent near-infrared (NIR) small molecule fluorescence probes for visualizing biological processes or diagnosing diseases. Herein, through the stepwise modulating structure of the silicon-rhodamine fluorophore (SR), we report three viscosity probes with allyl or methyl group as rotors, named SR-T-Al , SR-S-Al , and SR-T-Me . Among them, SR-T-Al demonstrates better viscosity responsibility from 1.0 to 1410.4 cP of viscosity. Therefore, the probe of SR-T-Al is successfully applied to sensitively monitor lysosome microscopic viscosity changes of living cells induced by oxygen stress. What's more, based on its advantages in NIR emission (669 nm) and large Stokes shift (201 nm), we also use it to image variations of viscosity in an acute hepatitis mouse induced by carbon tetrachloride. Both time and concentration-dependent induction models display the great ability of SR-T-Al to detect viscosity alteration. All the experimental results indicated that this allyl-rotor-based NIR viscosity probe could provide a general platform to monitor abnormal physiological processes and diseases relating to viscosity., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Co-published by Nanjing University and American Chemical Society.)

Accurate detection of target analytes and generation of high-fidelity fluorescence signals are particularly critical in life sciences and clinical diagnostics. However, the majority of current NIR-I fluorescent probes are vulnerable to pH effects resulting in signal distortion. In this work, a series of fluorescence-tunable and pH-independent probes are reported by combining optically tunable groups of unsymmetric Si-rhodamines and introducing the methoxy instead of the spiro ring on the benzene ring at position 9. To validate the concept, the leucine aminopeptidase response site was introduced into Si-2,6OMe-NH 2 with the best optical properties to synthesize Si-LAP for monitoring the intrahepatic LAP in vivo. Therefore, the design approach may provide a new and practical strategy for designing innovative functional fluorescent probes and generating high-stability and high-fidelity fluorescent signals.

Aberrant lysosomal alkalization is associated with various biological processes, such as oxidative stress, cell apoptosis, ferroptosis, etc. Herein, we developed a novel aminofluorene-based fluorescence probe named FAN to monitor the lysosomal alkalization-related biological processes by its migration from lysosome to nucleus. FAN possessed NIR emission, large Stokes shift, high pH stability, and high photostability, making it suitable for real-time and long-term bioimaging. As a lysosomotropic molecule, FAN can accumulate in lysosomes first and then migrate to the nucleus by right of its binding capability to DNA after lysosomal alkalization. In this manner, FAN was successfully used to monitor these physiological processes which triggered lysosomal alkalization in living cells, including oxidative stress, cell apoptosis, and ferroptosis. More importantly, at higher concentrations, FAN could also serve as a stable nucleus dye for the fluorescence imaging of the nucleus in living cells and tissues. This novel multifunctional fluorescence probe shows great promise for application in lysosomal alkalization-related visual research and nucleus imaging.

An "AND" logic gate-based NIR fluorescent probe Si-NH2-Glu was developed based on novel meso -amine Si-Rhodamine, which combined γ-glutamyl transpeptidase and pH dual-responsive sites. The features of Si-NH2-Glu enable it to be applied in orthotopic tumor imaging and fluorescence-guided surgery.

Background: Trimethylamine- N -Oxide (TMAO) is a proatherogenic and prothrombotic metabolite. Our study examined the association of plasma TMAO level with cardiovascular and all-cause mortality in hemodialysis (HD) patients., Methods: Patients who were at least 18 years-old and received HD for at least 6 months were enrolled within 6 months. Patients with coronary heart disease, congestive heart failure, arrhythmia, or stroke within 3 months before study onset were excluded. The primary endpoints were cardiovascular and all-cause death, and the secondary endpoint was cerebrovascular death., Results: We recruited 252 patients and divided them into a high-TMAO group (>4.73 μg/mL) and a low-TMAO group (≤4.73 μg/mL). The median follow-up time was 73.4 months (interquartile range: 42.9, 108). A total of 123 patients died, 39 from cardiovascular disease, 19 from cerebrovascular disease, and 65 from other causes. Kaplan-Meier analysis indicated that the high-TMAO group had a greater incidence of cardiovascular death (Log-Rank: p  = 0.006) and all-cause death (Log-Rank: p  < 0.001). Cox regression analysis showed that high TMAO level was significantly associated with cardiovascular and all-cause mortality. After adjustment for confounding, this association remained significant for cardiovascular mortality (TMAO as a continuous variable: HR: 1.18, 95%CI: 1.07, 1.294, p  < 0.001; TMAO as a dichotomous variable: HR: 3.44, 95%CI: 1.68, 7.08, p  < 0.001) and all-cause mortality (TMAO as a continuous variable: HR: 1.14, 95%CI: 1.08, 1.21, p  < 0.001; TMAO as a dichotomous variable: HR: 2.54, 95%CI: 1.71, 3.76, p  < 0.001)., Conclusions: High plasma TMAO level is significantly and independently associated with cardiovascular and all-cause mortality in HD patients.

Indoxyl sulfate (IS), a typical uremic toxin, is of great importance in the development of chronic kidney disease. In addition to its nephrotoxicity, previous studies have provided increasing evidence for its cardiovascular toxicity. The mechanism underlying IS‑induced cardiovascular toxicity has been elusive to date. The present study aimed to evaluate whether IS treatment could induce apoptosis of H9C2 cells, and used the endoplasmic reticulum (ER) stress‑modulator 4‑phenylbutyric acid (4‑PBA) to evaluate whether IS‑induced apoptosis is indeed associated with ERS. To evaluate whether IS induces apoptosis in H9C2 cardiomyocytes, cells were exposed to increasing concentrations of IS (500, 1,000, and 2,000 µM) for 24 h, and apoptosis was detected by flow cytometry. To determine whether IS‑induced apoptosis is associated with ERS, cells were divided into 4 groups: control group, PBA group, IS group and PBA+IS group. IS dose‑dependently induced apoptosis, and increased the expression of ER chaperones in H9C2 cells. Additionally, 4‑PBA treatment decreased IS‑induced apoptosis, and reduced ERS‑associated protein expression induced by IS. Therefore, the mechanism may be associated with the CCAAT‑enhancer‑binding protein homologous protein and c‑Jun N‑terminal kinase signaling pathways.

Two new guaiane-type sesquiterpenes, kwangsiensis A and B (1-2) were isolated from the roots of Curcuma kwangsiensis. Their structures were elucidated by extensive spectroscopic methods, including NMR, circular dichroism (CD) and high-resolution mass-spectrometry. The anti-inflammatory activity of the two compounds was evaluated on the basis of their inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in mouse RAW 264.7 macrophages. Compounds 1 and 2 showed moderate inhibitory activities with IC 50 values of 27.4 and 35.1 μM, respectively.

Objective: To investigate the effects of multi-day low dose ketamine infusion for postmastectomy pain syndrome (PMPS) after breast cancer surgery. Methods: This study was a prospective randomized controlled trial. From June 2015 to May 2016 in Affiliated Yiwu Hospital of Wenzhou Medical University, 66 patients with breast cancer surgery were randomly divided into control group (group C) and ketamine group (group K). Patients in group K were infused with 0.5 mg/kg of ketamine mixed in 250 ml of 0.9% normal saline in 1 h daily for 7 days. Patients in group C were infused the same dose of 0.9% normal saline. Anesthesia induction in both groups were given intravenous midazolam, sufentanil, propofol, vecuronium and intermittent positive pressure ventilation after tracheal intubation, anesthesia was maintained with propofol and remifentanil. After awakening, all patients were monitored in postanesthesia care unit (PACU) and given patient-controlled intravenous analgesia(PCIA). Pain scores were assessed using visual analogue scales (VAS) during PACU, 4 h, 24 h and 2-5 d after surgery, simultaneously analgesic requirement were recorded. Patients were evaluated Hospital Anxiety and Depression Scale (HADS) 5 d after surgery . The patients were followed up for 6 months. At 3 m, 6 m after surgery, the incidence of PMPS, the level of pain, pain site and HADS scale were assessed. Results: The VAS score uring PACU, 4 h, 24 h and 2-5 d after surgery in group K( (2.5±0.8), (2.4±0.5), (2.4±0.5), (2.0±0.4), (1.5±0.5), (1.0±0.4), 1(1), respectively) was lower than those in group C ((2.9±1.0), (2.9±0.6), (2.6±0.5), (2.3±0.5), (1.8±0.6), (1.5±0.5), 1(0), respectively). There was statistically difference between the two groups (all P <0.05). The consumption of analgesics required at each time postoperation in group K were also lower than that of group C(all P <0.05). Followed up for 6 months, 2 lost in group C, 1 lost in group K. The incidence of PMPS in group K at 3 months and 6 months after surgery was significantly lower(25% and 22%) than that in group C(52% and 45%)(χ(2)=4.729, 3.842, all P <0.05). There were no significant difference in pain level and site between two groups of PMPS patients (all P >0.05). There were no significant difference of HADS scale preoperative and 5 d after surgery between two groups (all P >0.05); and HADS scale in group K at 3 m and 6 m after surgery was significantly lower than that in group C(all P <0.05). Conclusion: Perioperative continuous multi-day low dose ketamine infusion can effectively reduce the incidence of PMPS after breast cancer surgery.

Background: It is widely accepted that chronic renal failure is associated with severe alterations of immune system. However, few studies looked into the immune alteration in earlier stage of chronic kidney disease (CKD) patients. To characterize immune defect in CKD patients, we performed lymphocyte subset analysis and explored its relationship to renal function in this population., Methods: 472 CKD patients were enrolled in this study. Lymphocyte subsets (CD19(+), CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD56(+)CD16(+)) were determined by flow cytometry. Clinical and laboratory data were collected. Patterns of immune cells in different stages of CKD were compared. Multivariate linear regression was used to evaluate the relationship between lymphocyte subset group and renal function. Correlation analysis was used to assess the relationship between lymphocyte subset and other clinical and laboratory data., Results: Decreased lymphocyte counts occurred long before the end stage of renal disease. Increased NK cell percentage was negatively related to estimated glomerular filtration rate (eGFR) (r = -0.259, p < 0.001) while B cell percentage was positively related to eGFR (r = 0.249, p < 0.001). Further multivariate linear regression showed increased B cell percentage (β = 16.470, 95%CI [1.018-31.922], p = 0.037) and decreased NK cell percentage (β = -10.659, 95%CI [-20.063 to -1.254], p = 0.026) were independently correlated with higher eGFR, respectively. Patients with lower NK cell percentage and higher B cell percentage tended to have the best renal function., Conclusions: Lymphocyte depletion and subset alteration occurred during the progress of CKD. Further studies are needed to clarify the role of immune system in CKD and to expand our knowledge about the effect of uremia on the structure and function of immune system.

Background and Objectives: Indoxyl sulfate, a protein-bound uremic toxin, may be associated with cardiovascular events and mortality in patients with CKD. This study aimed to investigate the relationship between indoxyl sulfate and heart failure in patients on hemodialysis., Design, Setting, Participants, & Measurements: Patients on hemodialysis for >6 months were enrolled within 6 months. Patients with congestive heart failure, angina pectoris, acute myocardial infarction, cerebral infarction, or cerebral hemorrhage within 3 months before the study or those <18 years old were excluded. The primary end point was first heart failure event during follow-up., Results: In total, 258 patients (145 men) with a mean age of 57.0 ± 14.6 years old were enrolled. Median plasma indoxyl sulfate level was used to categorize patients into two groups: the low-indoxyl sulfate group (indoxyl sulfate ≤ 2.35 μg/ml) and the high-indoxyl sulfate group (indoxyl sulfate >32.35 μg/ml). Then, patients were prospectively followed up for a median of 48.0 (interquartile range: 33.5-48.0) months. During follow-up, 68 patients experienced episodes of first heart failure. Kaplan-Meier analysis revealed the incidence of first heart failure event in the high-indoxyl sulfate group was significantly higher than in the low-indoxyl sulfate group (log rank P<0.001). Cox regression analysis showed indoxyl sulfate was significantly associated with first heart failure event (indoxyl sulfate as the continuous variable: hazard ratio, 1.02; 95% confidence interval [95% CI], 1.01 to 1.03; P=0.001; indoxyl sulfate as the dichotomous variable: hazard ratio, 3.49; 95% CI, 1.97 to 6.20; P<0.001). After adjustment for other confounding factors, the results remained significant (indoxyl sulfate as the continuous variable: hazard ratio, 1.04; 95% CI, 1.02 to 1.06; P<0.001; indoxyl sulfate as the dichotomous variable: hazard ratio, 5.31; 95% CI, 2.43 to 11.58; P<0.001)., Conclusions: Plasma indoxyl sulfate was associated with first heart failure event in patients on hemodialysis. Whether indoxyl sulfate is only a biomarker or involved in the pathogenesis of heart failure in hemodialysis warrants additional study., (Copyright © 2015 by the American Society of Nephrology.)

Beta-B2-crystallin (CRYBB2), encoded by Crybb2 gene, is a major protein in the mammalian eye lens that plays an important role in maintaining the transparency of the ocular lens. However, CRYBB2 also plays important roles in many extra-lenticular tissues and organs such as the retina, brain and testis. Our previous studies demonstrated that male Crybb2 deficient (Crybb2(-/-)) mice have reduced fertility compared with wild-type (WT) mice, while female Crybb2(-/-) mice exhibited reduced ovary weights and shorter estrous cycle percentages. Here we specifically investigated the role of CRYBB2 in the female reproductive system. Our studies revealed that ovaries from female Crybb2(-/-) mice exhibited significantly reduced numbers of primordial, secondary and pre-ovulatory follicles when compared with WT mice, while the rate of atretic follicles was also increased. Additionally, fewer eggs were collected from the oviduct of Crybb2(-/-) female mice after superovulation. Estrogen levels were higher in the metestrus and diestrus cycles of female Crybb2(-/-) mice, while progesterone levels were lower in diestrus cycles. Furthermore, the expression of survival and cell cycle genes, Bcl-2, Cdk4 and Ccnd2, were significantly decreased in granulosa cells isolated from female Crybb2(-/-) mice, consistent with the predominant expression of CRYBB2 in ovarian granulosa cells. Our results reveal a critical role for CRYBB2 in female fertility and specific effects on the proliferation and survival status of ovarian granulosa cells., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Diabetic retinopathy is a leading cause of reduced visual acuity and acquired blindness. Diabetes is known to alter the amount of retinal expression of the water-selective channels aquaporin 4 (AQP4). However, the function and impact of AQP4 in diabetic retinopathy is not well understood. In the present work, diabetes was induced by intraperitoneal injection of streptozotocin in Sprague-Dawley rats. Two weeks later, AQP4 shRNA (r) lentiviral particles or negative lentiviral particles were delivered by intravitreal injection to the eyes. Gene delivery was confirmed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and Western blotting analysis. Eight weeks later, BRB breakdown was measured using Evans blue dye. Images of retinal sections were obtained and the thicknesses of the retinas were determined. Retinal leukostasis measurement was performed using acridine orange leukocyte fluorography. The mRNA levels of IL-1β, IL-6, intercellular adhesion molecule 1 (ICAM-1), glial fibrillary acidic protein (GFAP) and vascular endothelial growth factor (VEGF) were determined using qRT-PCR method. AQP4 shRNA (r) lentiviral particles or negative lentiviral particles were transfected into rMC-1 cells to investigate its effect on inflammation induced by high glucose. Incubation with IL-1β or IL-6 was performed to test their effect on AQP4 expression in rMC-1 cells. In the current work, it was found that AQP4 expression was enhanced in the retina of diabetic rats. AQP4 knockdown led to exacerbation of retinopathy including enhancing retinal vascular permeability, retinal thickness, pro-inflammatory factors expression, and VEGF and GFAP expression in retinas of diabetic rats. AQP4 knockdown enhanced the expression of pro-inflammatory cytokines induced by high glucose in rMC-1 cells. In addition, AQP4 knockdown enhanced the release of IL-6 and VEGF from rMC-1 cells into the medium. Moreover, it was found that incubation with IL-1β or IL-6 suppressed AQP4 expression in rMC-1 cells. These results suggested that streptozotocin injection induced diabetes resulted in compensatory increases of AQP4 expression, and downregulation of AQP4 exacerbated diabetic retinopathy through aggravating inflammatory response, at last in part. Therefore, regulation of retinal function by AQP4 may attenuate diabetic retinopathy, offering a promising therapeutic strategy for diabetic retinopathy., (Copyright © 2012. Published by Elsevier Ltd.)

Aim: To explore the pathogenesis, influencing factors, ways of medical intervention and evaluation indicators of cataract by observing changes in serum biochemical indices in mice with targeted disruption of βB2-crystallin., Methods: Nine 6-week-old male mice with targeted knockout of βB2-crystallin were used as the study group, and nine age- and sex-matched normal wild-type mice as the control group. The genetype of the modeled mice was identified by PCR technique. Tropicamide and phenylephrine eye drops were used as the cycloplegic agents to observe changes in lens opacity with a slit-lamp. The lens was then removed and blood was collected for biochemical evaluation in the serum., Results: Two genotypes were successfully identified by PCR technique. Slit-lamp observation showed that the lens cortex was opaque and GSH level in the lens cortex was remarkably decreased in mice with βB2-crystallin deficiency compared with the control group (P<0.01). Serum Na(+), Cl(-), Ca(2+), Mg(2+) and Fe(2+) levels, ALT and AST activities, and TP, ALP, Cr, TC, GLU content were decreased significantly compared with the control group (P<0.05). There was no difference in LDH, P, Cu(2+), K(+) levels between the two groups (P>0.05)., Conclusion: Compared with the wild-type mice, serum biochemical indices underwent significant changes in mice with targeted disruption of βB2-crystallin gene, especially with abnormal distribution of Na(+)&Ca(2+), which induced the formation of cataract.