Oxidative stress biomarker triggered multiplexed tool for auxiliary diagnosis of atherosclerosis.

Oxidative stress is integral in the development of atherosclerosis, but knowledge of how oxidative stress affects atherosclerosis remains insufficient. Here, we design a multiplexed diagnostic tool that includes two functions (photoacoustic imaging and urinalysis), for assessing intraplaque and urinary malondialdehyde (MDA), a well-recognized end-product of oxidative stress. Molecular design is conducted to develop the first near-infrared MDA-responsive molecule (MRM). Acid-unlocked ratiometric photoacoustic nanoprobe is designed to report intraplaque MDA, enabling it to reflect plaque burden. Furthermore, MRM is tailored for urinary MDA detection with excellent specificity in a blind study. Moreover, we found a significant difference in urinary MDA between healthy adults and atherosclerotic patients (more than 600 participants). Combining these two functions, such a multiplexed diagnostic tool can dynamically report intraplaque and systemic oxidative stress levels during atherosclerosis progression, pneumonia infection, and drug treatment in atherosclerotic mice, which is promising for the auxiliary diagnosis of atherosclerosis.

Engineering of cyanine-based nanoplatform with tunable response toward reactive species for ratiometric NIR-II fluorescent imaging in mice.

High-quality second near-infrared (NIR-II) nanoprobes are of great significance for real-time bioimaging and medical diagnosis. Cyanine is an important class of fluorophores to construct activatable probes; however, there are still significant challenges hindering their biological applications, including weak fluorescence in aqueous solution, instability, and insufficient specificity. Herein, an integrated engineering strategy is conducted to develop the cyanine-based activatable NIR-II nanoplatforms with bright, stable emission and high specificity. Specifically, poly(styrene-co-maleic anhydride) (PSMA) is employed to encapsulate NIR-II fluorescent molecules (IR1048) to render the stable and bright NIR-II nanoparticles (PSMA@IR1048 NPs). By charge-modulated strategy, a series of cyanine-fluorophores are loaded on the surface of PSMA@IR1048 NPs and exhibit tunable response toward reactive species. Combing those two strategies, NIR-II ratiometric fluorescent nanoprobes (RNPs, including RNP1, RNP2, and RNP3) are constructed; among them, RNP2 displays hypochlorous acid (HClO) responsive performance and generates a higher NIR-II fluorescent ratio (FL2/FL1) signal. Such nanoprobe can reliably report the pathological HClO level in models of diabetic liver injury and lower limb ischemia-reperfusion (I/R) injury mice. Our study paves an engineering strategy to construct cyanine-based stable, bright, and specific NIR-II probes for bioimaging.

Rational Design of a Double-Locked Photoacoustic Probe for Precise In Vivo Imaging of Cathepsin B in Atherosclerotic Plaques.

Atherosclerotic plaque rupture is a significant cause of acute cardiovascular events such as heart attack and stroke, triggered by the decomposition of fiber caps induced by cysteine cathepsin. However, the accurate measurement of cathepsin B (CTB) activity in plaques is challenging due to the low specificity and insufficient penetration depth of available atherosclerosis-associated cathepsin fluorescent probes, hampering reliable assessment of plaque vulnerability. To address these limitations, we added both lipophilic alkyl chain and hydrophilic CTB substrate to the hemicyanine scaffold to develop a lipid-unlocked CTB responsive probe (L-CRP) that uses lipids and CTB as two keys to unlock photoacoustic (PA) signals for measuring CTB activity in lipophilic environments. Such properties allow L-CRP for the reliable imaging of specific CTB activities in foam cells and atherosclerotic plaques while keeping in silence toward CTB in lipid-deficient environments, such as M1-type macrophages and LPS-induced inflammatory lesions. Moreover, the activatable PA signals of L-CRP exhibit a deeper tissue penetration ability (>1.0 cm) than current CTB probes based on near-infrared fluorescent imaging (∼0.3 cm), suitable for atherosclerosis imaging in living mice. In atherosclerotic mice, L-CRP dynamically reports intraplaque CTB levels, which is well-correlated with the plaque vulnerability characteristics such as fiber cap thickness, macrophage recruitment, and necrotic core size, thus enabling risk stratification of atherosclerotic mice complicated with pneumonia. Moreover, L-CRP successfully identifies atherosclerotic plaques in excised human artery tissues, promising for auxiliary diagnosis of plaque vulnerability in clinical application.

Increased risks of maxillary sinus mucosal thickening in Chinese patients with periapical lesions.

Objectives: This study aimed to evaluate the effects of factors related to periapical lesions (PALs) on sinus membrane thickening (SMT) in the Chinese population using cone-beam computed tomography (CBCT). Methods: In this retrospective study, CBCT images (n = 512) of maxillary sinuses of 446 patients were evaluated by two examiners for SMT and PALs, PAL size, and the distance between the maxillary sinus floor (MSF), and the PAL's edge/root apex. The data were analyzed using analysis of variance, the Kruskal-Wallis test, χ2-test, and logistic regression. Results: A binary logistic regression model showed that the prevalence and severity of SMT > 2 mm were significantly associated with older age (>60 years) (odds ratio [OR]: 4.03, 95% confidence interval [CI]): 2.24-7.72, P < 0.001], male sex (OR: 2.08, 95% CI: 1.21-3.56, P < 0.006), and PALs (OR: 6.89, 95% CI: 3.93-12.08, P < 0.001). The type of contact and penetration between the MSF and PALs or root apex showed a more significant relation with SMT > 2 mm than did distance after adjusting for confounding factors, including age and sex (PALs: OR = 10.17 and 14.57, P < 0.001; root apex: OR = 3.49 and 5.86, P < 0.001). Conclusions: The prevalence and severity of SMT were significantly associated with older age, male sex, PALs, PAL size, and the distance between the MSF and PALs/root apex. Therefore, communication between dental surgeons and an otolaryngology specialist is important for the timely diagnosis and treatment of SMT of dental origin.

Semiconducting Polymer Nanoparticles-Manganese Based Chemiluminescent Platform for Determining Total Antioxidant Capacity in Diabetic Mice.

The total antioxidant capacity (TAC) is a key indicator of the body's resistance to oxidative stress injury in diabetic patients. The measurement of TAC is important for effectively evaluating the redox state to prevent and control the occurrence of diabetes complications. However, there is a lack of a simple, convenient, and reliable method to detect the total antioxidant capacity in diabetes. Herein, we design a novel chemiluminescent platform based on semiconducting polymer nanoparticles-manganese (SPNs-MnVII) to detect the total antioxidant capacity of urine in diabetic mice. We synthesize semiconducting polymer nanoparticles with four different structures and discover the ability of MnVII to produce singlet oxygen (1O2) that is employed to excite thiophene-based SPNs (PFODBT) to emit near-infrared chemiluminescence. Notably, the chemiluminescent intensity has a good linear relationship with the concentration of MnVII (detection limit: 2.8 µM). Because antioxidants (e.g., glutathione or ascorbic acid) can react with MnVII, such a chemiluminescent tool of SPNs (PFODBT)-MnVII can detect the glutathione or ascorbic acid with a larger responsive range. Furthermore, the total antioxidant capacity of urine from mice is evaluated via SPNs (PFODBT)-MnVII, and there are statistically significant differences between diabetic and healthy mice. Thus, this new chemiluminescent platform of SPNs (PFODBT)-MnVII is convenient, efficient, and sensitive, which is promising for monitoring antioxidant therapy of diabetes.

Comparison of cytokine level changes in gingival crevicular fluid between the aligner and pendulum appliance during early molar distalization : A single-center, prospective, observational study.

PURPOSE: There is currently a lack of evidence pertaining to gingival crevicular fluid cytokine levels in patients treated with the pendulum appliance. This study aimed to compare changes in cytokine secretion levels in gingival crevicular fluid (GFC) between the aligner and pendulum appliance in patients undergoing molar distalization. METHODS: GFC samples were collected from 28 participants with asymmetrical dental class II malocclusion who were undergoing molar distalization using aligners or pendulum appliances. An enzyme-linked immunosorbent assay was used to detect cytokine secretion levels during asymmetrical molar distalization for up to 14 days. Periodontal health indices and tooth movements were also assessed. RESULTS: No significant difference was found for the distalization distance between the two appliances. The Silness and Loe Plaque Index and Lobene Modified Gingival Index increased in the pendulum group but not in the aligner group at 14 days. Interleukin-1ß and tumor necrosis factor­α were upregulated in both groups. In the pendulum group, receptor activator of nuclear factor kappa­Β ligand and osteoprotegerin secretion levels were significantly upregulated and downregulated, respectively; smaller changes in these two cytokines were observed in the aligner group. CONCLUSIONS: Pendulum appliances exert stronger forces than aligners, which cause more changes in the secretion of inflammatory mediators in young patients.

Effect of Mesioangular Impaction of the Mandibular Third Molars on the Adjacent Mandibular Second Molars: An Imaging Study.

OBJECTIVE: To investigate the effect of mesioangular impaction of the mandibular third molars on distal caries and distal bone loss (DBL) in the adjacent mandibular second molars to provide a basis for prophylactic impacted third molar extraction. METHODS: We retrospectively analyzed panoramic radiographs of 736 patients with mesioangularly impacted mandibular third molars at the Department of Stomatology, Shanghai Fifth People's Hospital. The chi-square test, the Kruskal-Wallis test, and logistic regression were performed to analyze the relationship of patient age, sex, mesial inclination angle, distal caries, and DBL in the mandibular second molars. RESULTS: Patient aged over 30 years and a mesial inclination angle > 30° were risk factors for distal caries in the mandibular second molars (p < 0.05). Patients aged over 20 years and a mesial inclination angle of 60°-90° were risk factors for DBL (p < 0.05). CONCLUSION: In patients with mesioangularly impacted mandibular third molars, age and the mesial inclination angle were positively correlated with the severity of distal caries and DBL in the adjacent mandibular second molars. These findings could guide dental practitioners to prevent complications of the adjacent molars during early prophylactic extraction of the mandibular third molars with mesioangular inclination.

The relationship between FOSB and SOCS3 gene polymorphisms and the susceptibility to periodontitis and osteopenia in the Chinese population.

The aim is to analyze the relationship and significance of the FOS, FOSB, Suppressors of cytokine signaling (SOCS), and hypoxia-inducible factor 1 (HIF1) gene loci and their polymorphisms with periodontitis and osteopenia in the Chinese population. In this case-control study, data on questionnaires, periodontal examination, bone mineral density, and FOS, FOSB, SOCS3, and HIF1 gene loci and their polymorphisms were obtained from 474 participants. The data were analyzed using the analysis of variance, Kruskal-Wallis test, χ2 test, and logistic regression. The incidence of osteopenia was significantly increased in patients with periodontitis compared to controls (58.6 vs. 34.4%, P < 0.001). Accordingly, the risk was increased 2.21-fold compared with controls (95% CI 2.09-4.95). Osteopenia patients had a significantly higher risk of periodontitis than patients with normal bone density (OR = 3.22, 95% CI 2.09-4.94). There were significant positive associations between FOSB and SOCS3 polymorphisms and periodontitis and osteopenia susceptibility. Individuals carrying the G/G genotype of the FOSB gene rs708905 locus had an increased risk of periodontitis (OR = 5.06, 95% CI 2.36-10.86) and osteopenia (OR = 3.26, 95% CI 1.34-7.96). Compared with the C/C genotype, the A/A genotype of the FOSB rs8105114 locus was associated with a significantly higher risk of periodontitis (OR = 2.14, 95% CI 1.02-4.53) and osteopenia (OR = 2.85, 95% CI 1.12-7.22). Compared with the A/A genotype, the risk of periodontitis in the G/G genotype of the SOCS3 rs7207782 locus was increased 3.10-fold (P < 0.001), and the risk of osteopenia was increased 2.01-fold (P = 0.023). There was a bidirectional relationship between periodontitis and osteopenia. The rs708905 G/G and rs8105114 A/A genotypes of FOSB and the rs7207782 G/G genotype of SOCS3 were risk factors for both periodontitis and osteopenia in the Chinese population, which could increase knowledge about disease­specific and cross­disease genetic pattern.

Effect of silencing of mediator of DNA damage checkpoint protein 1 on the growth of oral squamous cell carcinoma in vitro and in vivo.

Mediator of DNA damage checkpoint protein 1 (MDC1) is involved in DNA damage repair and has been linked to tumor invasion, metastasis, and prognosis. This study investigated the effects of MDC1 in oral squamous cell carcinoma (OSCC) in vitro and in vivo. RNA interference-mediated knockdown of MDC1 was performed in two OSCC cell lines (Tca-8113 and KB). Real-time PCR and western blotting were performed to determine expression of mRNA and protein, respectively, of MDC1. Cell viability was assessed using the MTT assay. Colony-formation assays were performed by staining with 0.5% crystal violet. Cell migration and invasion were detected by Transwell assays. The role of MDC1 in OSCC was examined in vivo via injection of Tca-8113 cells transfected with MDC1 small interfering (si)RNA or negative-control siRNA into a mouse xenograft model of OSCC. Our results showed that MDC1 knockdown decreased cell proliferation. Inhibition of MDC1 decreased colony formation of Tca-8113 and KB cells by 62% and 68%, respectively, and MDC1 knockdown reduced the number of migratory and invasive cells compared with the control group. Moreover, the xenograft mouse model of MDC1 knockdown showed reduced tumor growth. Our study suggests that MDC1 plays a role in tumorigenesis and might be a potential target for the treatment of patients with OSCC.

[The effect of different gutta-percha master cones on first penetration depth of spreader and apical microleakage].

PURPOSE: This study was to evaluate the effect of different master cones on apical microleakage through comparing first penetration depth of spreader and the leakage concentration of glucose in the curved root canal of the extracted posterior teeth filled with different gutta-percha master cones. METHODS: Sixty-six extracted roots of human posterior teeth with a single, curved canal were collected and divided into five experimental groups of 11 roots in each group (Group A:master cone 0.06 tapered, 25#; Group B: 0.06 tapered, 20#; Group C: 0.04 tapered, 30#; Group D: 0.04 tapered, 25#; Group E: 0.02 tapered, 25#) using cold lateral condensation as positive control group(group F, AH-PLUS sealer) of 5 roots and negative control (group G, integrated teeth) of 6 roots. The first penetration depth of spreader and the concentration of glucose in apical reservoir at 1, 2, 4, 7, 10, 15, 20 and 30 days were measured, respectively. The data was analysed with ANOVA and LSD test using SPSS 16.0 software package. RESULTS: The value of working length minus the first penetration depth of spreader in group D was significantly smaller than that in group A and group C(P<0.05),and that in group E was significantly smaller than that in group A ,B,C(P<0.05), but no significant difference(P>0.05) in other groups. The concentration of leakage glucose in group F was significantly higher than that in the other six groups(P<0.05),and there were no significant difference in the five experimental groups (P>0.05),which had no significant difference from the negative control group G(P>0.05). CONCLUSIONS: Choosing smaller tapered master cone could get better apical filling in curved canals, but it has no superiority in apical sealing in short time.

[The effect of smear layer on apical microleakage].

PURPOSE: To evaluate the effect of smear layer on apical microleakage in extracted root-filled teeth by SEM and glucose penetration model. METHODS: 51 straight single-rooted human teeth were randomly divided into 5 groups, instrumented and irrigated differently by groups. 11 teeth in group A were irrigated with 17% EDTA and 1% NaClO,11 teeth in group B with 1% tetracycline hydrochloride and 1% NaClO,11 teeth in group C with 10% citric acid and 1% NaClO, 9 teeth in group D with 1% NaClO only and 9 teeth in group E with 0.9% NaCl. One tooth in each group was observed under scanning electron microscopy. The other teeth were obturated using lateral condensation technique. Leakage along the root filling was measured by the concentration of glucose in apical reservoir at 1, 2, 4, 7, 10, 15, 20 and 30 days respectively with glucose oxidase method. The data were analyzed with repeated measure ANOVA and SNK-q test using SPSS 16.0 software package. RESULTS: No smear layer on the root canal wall was observed in group A, B and C, the leakage was lower than that in group D and E (P<0.05). The smear layer occurred in quantity in group D, E and their leakage increased. The microleakage was higher in group E than that in group D (P<0.05). CONCLUSION: Removal of smear layer on the root canal wall could reduce the root apical microleakage.

[Experimental study of preparing curved root canals with different instrument].

OBJECTIVE: To compare the shaping ability and the influence on apical foramen among hand ProTaper, stainless steel K-files and rotary ProTaper in preparing different curved root canal. METHODS: Forty simulated resin root canal blocks were randomly divided into four groups and prepared by hand ProTaper, stainless steel K-file and rotary ProTaper, respectively. Of them, 12 blocks in group A, B, C consist of six 200 curved root canals and six 30 degrees curved root canals each group. The curvature of the other 4 blocks in group D was less than 5 degrees. Taking photos of the models to the root canal orthotopically and apical foramen using digital camera before and after instrumentation. Finally, the transportation of root canal and the size of apical foramen were analyzed using special image software Auto-CAD. RESULTS: The transportation of center in group B was the highest than that in group A and group C (P<0.05). In some portions of root canal, the transportation of center in group C was higher than that in group A. The size of the apical foramen in group B was significantly bigger than the other groups and the size of the apical foramen in 30 degrees root canal was significantly bigger than that in 20 degrees root canal after instrumentation (P<0.05). There was no significantly different between group A and group C, though the size of apical foramen in group C was bigger than that in group A at the same curvature, and that in 30 degrees root canal was bigger than in 20 degrees root canal (P>0.05). CONCLUSION: Both of the two instruments engender root canal transportation, and curvature is the main reason of transportation. Comparing with stainless steel K-files, NiTi files can maintain the shape of the root canal and apical foramen well.