Staining Index Research Articles

Comparative Evaluation of Efficacy of Chitosan and Chlorhexidine Mouthwash in Plaque Control and Gingivitis: An Observational Study.

Mouth rinse is a highly effective chemical solution for managing plaque and gingivitis. This study evaluated and compared the efficacy of chitosan (CH)-rich mouthwash with standard chlorhexidine (CHX) mouthwash in reducing gingivitis, plaque, halitosis, and stains. This retrospective, observational study was conducted on the clinical case records of 60 patients who were treated with CH mouthwash (formulated and dispensed from the department) as group 1 and CHX mouthwashas group 2 for at least three months. Indices such as the plaque index (PI), gingival index (GI), stain index (SI), and halitosis index (HI) were obtained at baseline (T0) and the end of three months of mouthwash use (T1) from the case records. The data were subjected to statistical analysis. Both groups effectively reduced all indices from T0 to T1; however, group 1 was more effective than group 2 in reducing gingival, plaque, and stain scores. The results were statistically significant (p<0.05) for all indices, and both groups were equally effective in reducing halitosis. Male sex showed a significant positive correlation with PI, GI, and SI, whereas female sex showed a positive correlation with PI and HI. Based on the findings of the present study, it was concluded that the CH mouthwash was more effective in reducing plaque, gingival, and staining scores than the CHX mouthwash. Both types of mouthwash were equally effective in reducing halitosis scores.

Assessing the therapeutic role of trehalose and hyaluronic acid: implications for patient care.

This Systematic review aims to assess the efficacy of trehalose and hyaluronic acid in enhancing ocular recovery post-cataract surgery, focusing on their impact on tear film stability, ocular surface integrity, and patient-reported outcomes. A comprehensive search was conducted across MEDLINE, PubMed, and Cochrane Library databases to identify randomized controlled trials investigating the efficacy of trehalose, hyaluronic acid, or their combination in post-cataract surgery care. The inclusion criteria focused on peer-reviewed studies in English, detailing outcomes relevant to ocular recovery such as tear film stability, ocular surface integrity, patient-reported discomfort, or visual acuity (VA). The quality of the included studies was assessed using the Cochrane Risk of Bias Tool and synthesized the data qualitatively. Four qualitative investigations met the inclusion criteria. The studies collectively assessed the efficacy of a 3% trehalose and 0.15% hyaluronic acid eye drop solution in reducing postoperative eye symptoms compared to various control solutions. Parameters measured included tear break-up time (TBUT), Fluorescein staining, tear production (Schirmer test), and Ocular Surface Disease Index (OSDI) scores. The results indicated significant improvements in tear film stability and ocular surface health for the treatment groups compared to controls, with a notable decrease in patient-reported discomfort. The study showed an improvement of - 18 (± 14.6) in the treatment group compared to - 7 (± 8.0) in the control group for OSDI. For TBUT, the treatment group improved by 3 (± 1.2) s, whereas the control group improved by 0.3 (± 0.71) s. VA, measured on a scale of 0-100, increased to 17 (± 0.7) in the treatment group compared to 15 (± 1.1) in the control group. Trehalose and hyaluronic acid may be beneficial in the postoperative period by enhancing tear film stability and ocular surface health. While the results are promising, further research is needed to confirm these findings, understand the mechanisms of action, and explore broader applications.

Clinical Comparison of Stain Removal Efficacy between Glycine Powder Air Polishing System and Ultrasonic Scaling in Removing Dental Plaque and Stains: A Split-mouth Randomized Clinical Trial

Abstract Introduction: Glycine powder air polishing (GPAP) has emerged as a less invasive option for periodontal maintenance therapy. This study aims at comparing the efficacy of GPAP in removing plaque and stains as compared to conventional ultrasonic scaling (USS). Materials and Methods: The present split-mouth randomised clinical trial recruited 20 adults with healthy gingiva or mild gingivitis. Removal of plaque and stains was performed by USS in one maxillary quadrant and by GPAP in the other. Clinical parameters assessed included stain scores by Lobene Stain Index, plaque level by Silness-Loe plaque Index, and pain scores using the Visual Analog Scale. The assessments were performed before and after the treatment and at a 15-day follow-up visit. Results: Significantly lower (P &lt; 0.05) levels of plaque and stains were achieved by both interventions as compared to those before the treatment. While immediate postoperative plaque levels were significantly (P &lt; 0.05) lower following USS, the difference failed to reach statistical significance (P &gt; 0.05) 1 week after the treatment. On the contrary, the stain scores were significantly lower after treatment and at the 7th-day follow-up after GPAP as compared to USS. A significantly greater number of patients (P &lt; 0.05) reported pain to be absent or only mild following GPAP, while those treated with USS experienced pain of severe intensity. Conclusion: Both GPAP and USS are effective in stain and plaque removal. However, GPAP achieved superior stain removal outcomes and was associated with significantly lower pain scores, suggesting it was more comfortable for patients compared to USS.

Hydrogen sulfide alleviates cadmium stress in germinating carrot seeds by promoting the accumulation of proline

Carrot (Daucus carota L.), a widely cultivated economically vegetable from the Apiaceae family, is grown globally. However, carrots can be adversely impacted by cadmium (Cd) pollution in the soil due to its propensity to accumulate in the fleshy root, thus impeding carrot growth and posing health hazards to consumers. Given the potential of hydrogen sulfide (H2S) to improve plant resistance against Cd stress, we treated germinating carrot seeds with varying concentrations of sodium hydrosulfide (NaHS), aiming to alleviate the toxic impacts of Cd stress on carrot seed germination. The results revealed that carrot seeds treated with a concentration of 0.25 mM NaHS displayed better seed germination-associated characteristics compared to seeds treated with NaHS concentrations of 0.1 mM and 0.5 mM. Further investigation revealed a rise in the expression levels of L-cysteine desulfhydrase and D-cysteine desulfhydrase, along with enhanced activity of L-cysteine desulfhydrase and D-cysteine desulfhydrase among the NaHS treatment group, thereby leading to H2S accumulation. Moreover, NaHS treatment triggered the expression of pyrroline-5-carboxylate synthase and pyrroline-5-carboxylate reductase and promoted the accumulation of endogenous proline, while the contents of soluble sugar and soluble protein increased correspondingly. Interestingly, since the application of exogenous proline did not influence the accumulation of endogenous H2S, suggesting that H2S served as the upstream regulator of proline. Histochemical staining and biochemical indices revealed that NaHS treatment led to elevated antioxidant enzyme activity, alongside a suppression of superoxide anion and hydrogen peroxide generation. Furthermore, high performance liquid chromatography analysis revealed that NaHS treatment reduced Cd2+ uptake, thereby promoting germination rate, seed vitality, and hypocotyl length of carrot seeds under Cd stress. Overall, our findings shed light on the application of NaHS to enhance carrot resistance against Cd stress and lay a foundation for exploring the regulatory role of H2S in plants responding to Cd stress.

Competitive swimming and dental staining among Australian children.

Swimming has classically been associated with a distinctive type of extrinsic dental staining. However, its prevalence and characteristics have not been explored in Australia. This case controlled cross-sectional cohort study aimed to determine the prevalence and severity of dental staining among children who swim competitively compared to non-swimmers and its impact on oral health-related quality of life (OHRQL). Parents and children from Western Australian swimming clubs completed a survey assessing swimming routine, risk factors for staining and OHRQL. Intra-oral photographs were taken and staining of anterior teeth was assessed using a modified Lobene Stain Index. A Global Stain Score (GSS) was calculated and compared against age-matched non-swimmer controls. Maxillary anterior teeth from swimmers (n = 400) and non-swimmers (n = 400) were analysed. Staining prevalence was greater in swimmers (83%) than non-swimmers (44%). GSS scores were positively correlated with both the total lifetime swimming hours and practice time (P < 0.05), with a significant difference in GSS between swimmers and non-swimmers (P = <0.001, OR: 6.21). Dietary factors for staining were not significantly correlated with GSS. Swimmers had a greater risk of developing staining than non-swimmers, and this negatively impacted their OHRQL. The extent and intensity of staining were associated with the amount of swimming.

Optimal treatment strategy for low-grade spinal cord astrocytoma: a retrospective, multicenter analysis by the Neurospinal Society of Japan

OBJECTIVE Primary spinal cord gliomas are rare, and among these astrocytomas (WHO grade II) are much rarer. The optimal treatment strategy thus remains unclear. The authors conducted a multicenter study led by the Neurospinal Society of Japan (NSJ) to analyze treatment policies and outcomes. The aim was to present optimal treatment methods for spinal cord astrocytoma and to identify predictors of better outcomes. METHODS Among 1033 consecutive cases of spinal cord intramedullary tumors treated surgically at 58 centers affiliated with the NSJ, 57 patients were diagnosed with diffuse astrocytoma (WHO grade II) and were enrolled in the present study. Among these 57 patients, treatment methods, outcomes, and tumor proliferation rate as evaluated by the MIB-1 staining index (SI) were analyzed, and the optimal treatment method for spinal cord astrocytomas (grade II) was determined. In addition, the authors searched for factors predicting better treatment outcomes. RESULTS Treatment for spinal cord astrocytoma comprised three methods: surgery alone in 30 patients, adjuvant radiation therapy in 13 patients, and adjuvant chemoradiotherapy in 13 patients. One patient who did not undergo surgery was excluded from survival analysis. Treatment with surgery alone or surgery with radiotherapy was associated with significantly longer overall and progression-free survivals than that with adjuvant chemoradiotherapy. Patients treated with radiation therapy had a higher MIB-1 SI than those treated with surgery alone. The extent of tumor resection tended to correlate with longer survival. In contrast, postoperative neurological worsening showed the inverse order. Adjuvant chemoradiotherapy was associated with the shortest survival in both total cases and recurrent cases. The optimal cutoff value of MIB-1 SI for predicting longer survival by surgery alone was less than 4.0%. CONCLUSIONS The optimal treatment for spinal cord astrocytoma is maximal tumor resection without neurological impairment. When some tumor remains in patients with an MIB-1 SI less than 4.0%, a wait-and-see approach is optimal. If the MIB-1 SI is higher than 4.0%, local radiation therapy is recommended. Adjuvant chemotherapy is not recommended for the treatment of grade II spinal cord astrocytoma.

Expression patterns of mismatch repair proteins in cervical cancer uncover independent prognostic value of MSH-2

ObjectiveAlthough early-detected cervical cancer is associated with good survival, the prognosis for late-stage disease is poor and treatment options are sparse. Mismatch repair deficiency (MMR-D) has surfaced as a predictor...

Paeonol ameliorates hyperlipidemia and autophagy in mice by regulating Nrf2 and AMPK/mTOR pathways

BackgroundHyperlipidemia, inadequate diet, and excessive medication increase the risk of cardiovascular disease. Paeonl (Pae), a phenolic compound found in Peony and Angelica dahurica, can alleviate lipid metabolism disorders and lipotoxicity. However, the molecular mechanism of Pae alleviating hyperlipidemia remains unclear and needs to be further explored. PurposeIn this study, we explored whether Pae can prevent hyperlipidemia and investigated the molecular mechanisms. MethodsThe effects of Pae (30, 45, 60mg·kg−1) on hyperlipidemia in Tyloapol-induced WT mice and Nrf2 knockout mice (Pae: 60mg·kg−1) were detected by oil red O staining, HE staining, TG, TC and other indexes. The expression levels of proinflammatory mediators, key lipid proteins and autophagy signaling pathway proteins were analyzed by enzyme-linked immunosorbent assay, western blot and immunofluorescence. The molecular mechanism of Pae alleviating hyperlipidemia was explored through molecular docking technique and in vivo and in vitro experiments. ResultsSeveral studies indicated that Pae effectively improved tyloxapol (Ty)-induced lipid metabolism disorder, as evidenced by decreased triglyceride content, increased carnitine palmitoyltransferase 1 (CPT1), and Sirtuin 1 (Sirt1) protein expression. In addition, Pae ameliorated hyperlipidemia by activating the AMPK/ACC and PI3K/mTOR pathways. Interestingly, the therapeutic effect of Pae on hyperlipidemia was markedly reduced in Nrf2−/− mice. Molecular docking results indicated that Pae and Nrf2 exhibited good binding ability, suggesting that Nrf2 is a core target mediating the effects of Pae in the treatment of hyperlipidemia. Taken together, Pae alleviated hyperlipidemia in vivo and ameliorated lipid accumulation in vitro by activating AMPK/ACC and PI3K/mTOR signaling pathways via Nrf2 binding. ConclusionOur data suggest that paeonol can ameliorate hyperlipidemia and autophagy in mice by regulating Nrf2 and AMPK/mTOR pathways, and it has potential therapeutic value in the occurrence and development of hyperlipidemia.

DDR2 expression in breast cancer is associated with blood vessel invasion, basal-like tumors, tumor associated macrophages, regulatory T cells, detection mode and prognosis

Discoidin Domain Receptor 2 (DDR2) is a receptor tyrosine kinase for collagen, stimulating epithelial-mesenchymal transition and stiffness in breast cancer. Here, we investigated levels of DDR2 in breast tumor cells in relation to vascular invasion, TIL subsets, macrophages, molecular tumor subtypes, modes of detection and prognosis. This retrospective, population-based series of invasive breast carcinomas from the Norwegian Screening Program in Vestfold County (Norway), period 2004–2009, included 200 screening patients and 82 cases detected in screening intervals. DDR2 was examined on core needle biopsies using a semi-quantitative, immunohistochemical staining index and dichotomized as low or high DDR2 expression. Counts of macrophages and TIL subsets were dichotomized based on immunohistochemistry using TMA. We also recorded blood or lymphatic vessel invasion (BVI or LVI) as present or absent by immunohistochemistry. High expression of DDR2 in tumor cells showed significant relation with high counts of CD163+ macrophages (p < 0.001) and FOXP3 TILs (p = 0.011), presence of BVI (p = 0.028), high tumor cell proliferation by Ki67 (p = 0.033), ER negativity (p = 0.001), triple-negative cases (p = 0.038), basal-like features (p < 0.001) as well as interval detection (p < 0.001). By multivariate analysis, high DDR2 expression was related to reduced recurrence-free survival (HR, 2.3, p = 0.017), when examined together with histologic grading, lymph node assessment, tumor diameter, BVI, and molecular tumor subtype. This study supports a link between high DDR2 expression, high counts of macrophages by CD163 (tumor associated) and regulatory T cells by FOXP3 together with the presence of BVI, possibly indicating increased tumor motility and intravasation in aggressive breast tumors.

Oral Papillary Squamous Cell Carcinoma and Oral Squamous Cell Carcinoma: A Histopathological and Immunohistochemical Comparative Study

PurposeThe aim of the study is to investigate the immunohistochemical expression of both Alpha smooth muscle actin and Transforming Growth Factor beta and compare their expression in oral papillary squamous cell carcinoma with their expression in different histological grades of oral squamous cell carcinoma. A correlation between these immuno-histochemical expressions and histological findings will then be performed. The research question is “Do the percentages of α-SMA and TGF-β immune-expression in OPSCC differ from that in the conventional OSCC?”.MethodsThis will be achieved by collecting archival blocks of oral papillary squamous cell carcinoma and different grades of oral squamous cell carcinoma, staining the specimens with Transforming Growth Factor beta and alpha smooth muscle actin, then measuring the mean staining index of expression in each group and the area percent of both markers.ResultsResults revealed that transforming growth factor beta expression in the epithelium was high in all cases of well-differentiated squamous cell carcinoma, most oral papillary squamous cell carcinoma, and poorly differentiated oral squamous cell carcinoma. On the other hand, different grades of oral squamous cell carcinoma showed a high staining index of alpha smooth muscle actin expression in the stroma. While cases of oral papillary squamous cell carcinoma were either moderate or low-staining.ConclusionsOral papillary squamous cell carcinoma has a favourable prognosis compared to different histological grades, and the prognosis does not depend only on histological grade but also on other prognostic factors.

Expression of Forkhead Box M1 and Anticancer Effects of FOXM1 Inhibition in Epithelioid Sarcoma

Epithelioid sarcoma (ES) is a rare aggressive sarcoma that, unlike most soft-tissue sarcomas, shows a tendency toward local recurrence and lymph node metastasis. Novel antitumor agents are needed for ES patients. Forkhead box transcription factor 1 (FOXM1) is a member of the Forkhead transcription factor family and is associated with multiple oncogenic functions; FOXM1 is known to be overexpressed and correlated with pathogenesis in various malignancies. In this study, we immunohistochemically analyzed FOXM1 expression levels and their clinical, clinicopathologic, and prognostic significance in 38 ES specimens. In addition, to investigate potential correlations between FOXM1 downregulation and oncologic characteristics, we treated ES cell lines with thiostrepton, a naturally occurring antibiotic that inhibits both small interfering RNA (siRNA) and FOXM1. In the analyses using ES samples, all 38 specimens were diagnosed as positive for FOXM1 by immunohistochemistry. We separated specimens into high (n = 19) and low (n = 19) FOXM1–protein expression groups by staining index score, and into large (n = 12), small (n = 25), and unknown (n = 1) tumor-size groups using a cutoff of 5 cm maximum diameter. Although there were significantly more samples with high FOXM1 expression in the large tumor group (P = .013), there were no significant differences with respect to age (P = 1.00), sex (P = .51), primary site of origin (P = .74), histologic subtypes (P = 1.00), depth (P = .74), or survival rate (P = .288) between the high and low FOXM1–protein expression groups. In the in vitro experiments using ES cell lines, FOXM1 siRNA and thiostrepton successfully downregulated FOXM1 mRNA and protein expression. Furthermore, downregulation of FOXM1 inhibited cell proliferation, drug resistance against chemotherapeutic agents, migration, and invasion and caused cell cycle arrest in the ES cell lines. Finally, cDNA microarray analysis data showed that FOXM1 regulated cIAP2, which is one of the apoptosis inhibitors activated by the TNFα-mediated NF-κB pathway. In conclusion, the FOXM1 gene may be a promising therapeutic target for ES.

A proposed index of myocardial staining for vein of marshall ethanol infusion

Abstract Background Mitral isthmus(MI) conduction block is a fundamental step in anatomical approach treatment for persistent atrial fibrillation(PeAF). However, MI block is hardly achievable with endocardial ablation only. Retrograde ethanol infusion(EI) into the vein of Marshall(VOM) facilitates MI block. Fluorographic myocardial staining(MS) during VOM-EI could be helpful in predicting procedural alcoholization outcome even if its role is qualitatively assessed in the routine. Purpose The aim was to quantitatively assess MS during VOM-EI and to evaluate its association with MI block achievement. Methods Consecutive patients undergoing catheter ablation for PeAF at our Institution from February 2022 to May 2023 were considered. Patients with identifiable VOM were included. A proposed index of MS(MSI) was retrospectively calculated in each included patient. Correlation of MSI with low voltage zones(LVZ) extension after VOM-EI and its association with MI block achievement were assessed. Results 42 patients out of 49 (85.8%) had an identifiable VOM. MI block was successfully achieved in 35 patients out of 42 (83.3%). MSI was significantly associated with the occurrence of MI block (OR 1.24 (1.03 – 1.48); p = 0.022). A higher MSI resulted in reduced ablation time (p = 0.014) and reduced radiofrequency applications (p = 0.002) to obtain MI block. MSI was also associated with MI block obtained by endocardial ablation only (OR 1.07 (1.02 – 1.13); p = 0.002). MSI was highly correlated with newly formed LVZ extension (r = 0.776; p = 0.001). Conclusion In our study cohort, optimal MSI predicts MI block and facilitates its achievement with endocardial ablation only.Myocardial stainingSpline plots

Validation of an automated iron stain process for use with bone marrow aspirate smear slides.

The assessment of bone marrow iron stores is typically performed on an aspirate smear slide that has been manually stained by a technologist using a commercially available kit. This approach can contribute to inconsistent results and limit the broad use of iron staining in bone marrow specimens, particularly when laboratories have low staffing and/or high specimen volumes. Here, we describe the adaptation and validation of the Ventana Benchmark automated stainer and iron stain kit for routine clinical use of staining iron in bone marrow aspirate smear slides. We assessed accuracy and precision of the Ventana automated iron staining protocol compared to the Perls Prussian blue manual iron staining index method. Hematopathologists assigned Gale scores and enumerated the percentages of erythroid sideroblasts on paired patient bone marrow aspirate smear slides stained by the automated method and the manual iron staining method. We found a similar level of performance of the Ventana automated iron stain relative to the index manual method (as assessed by Pearson correlation and Bland-Altman analyses). In addition, there was low imprecision between replicates performed via the automated iron stain protocol. We also report superior qualitative findings of the automated method in ease of localization of iron storage, visualization of sideroblasts, and counterstain consistency. Automated iron staining of bone marrow aspirate smear slides performed similarly to the manual method and may allow for accurate routine evaluation of bone marrow iron stores as part of bone marrow analysis.

The anti-non-small cell lung cancer effect of Diosbulbin B: Targeting YY1 induced cell cycle arrest and apoptosis

BackgroundToxic components frequently exhibit unique characteristics and activities, offering ample opportunities for the advancement of anti-cancer medications. As the main hepatotoxic component of Dioscorea bulbifera L. (DB), Diosbulbin B (DIOB) has been widely studied for its anti-tumor activity at nontoxic doses. However, the effectiveness and mechanism of DIOB against non-small cell lung cancer (NSCLC) remains unclear. PurposeTo evaluate the anti-NSCLC activity of DIOB and to elucidate the specific mechanism of action. MethodThe effect of DIOB on NSCLCL in vitro was evaluated through CCK8, colony formation, and flow cytometry. The in vivo efficacy and safety of DIOB in treating NSCLC were assessed using various techniques, including HE staining, tunel staining, immunohistochemistry, and biochemical index detection. To understand the underlying mechanism, cell transfection, western blotting, molecular docking, cellular thermal shift assay (CESTA), and surface plasmon resonance (SPR) were employed for investigation. ResultsDIOB effectively hindered the progression of NSCLC both in vitro and in vivo settings at a no-observed-adverse-effect concentration (NOAEC) and a safe dosage. Specifically, DIOB induced significant G0/G1 phase arrest and apoptosis in A549, PC-9, and H1299 cells, while also notably inhibiting the growth of subcutaneous tumors in nude mice. Mechanistically, DIOB could directly interact with oncogene Yin Yang 1 (YY1) and inhibit its expression. The reduction in YY1 resulted in the triggering of the tumor suppressor P53, which induced cell cycle arrest and apoptosis in NSCLC cells by inhibiting the expression of Cyclin A2, B2, CDK1, CDK2, CDK4, BCL-2, and inducing the expression of BAX. In NSCLC cells, the induction of G0/G1 phase arrest and apoptosis by DIOB was effectively reversed when YY1 was overexpressed or P53 was knocked down. Importantly, we observed that DIOB exerted the same effect by directly influencing the expression of YY1-regulated c-Myc and BIM, particularly in the absence of P53. ConclusionFor the inaugural investigation, this research unveiled the anti-NSCLC impact of DIOB, alongside its fundamental mechanism. DIOB has demonstrated potential as a treatment agent for NSCLC due to its impressive efficacy in countering NSCLC.

Strawberry Gel with a Concentration of 83% and 75% as an Extrinsic Stain Remover

Strawberries have many benefits, where the malic acid contained in strawberries acts as a substance that will erode and remove several stains on the surface of teeth, including cleaning extrinsic stains. Gel preparations are widely used in medicines, cosmetics and food products, including toothpaste and teeth whitening agents. The purpose of this study was to determine the effectiveness of strawberry gel with a concentration of 83% and 75% in cleaning teeth with extrinsic stain on academic member of the Health Polytechnic Jakarta I. This study was an Experiment on 30 people who had upper and lower anterior teeth with extrinsic stain as samples. Data collection was done by applying strawberry gel on the surface of the anterior teeth, allowed to stand for 5 minutes, then brushed and rinsed with water. This procedures was done 2 times a day for 5 consecutive days. Changes in extrinsic stain are measured using the Lobene Stain Index which measures the intensity and area of the stain, as well as a combined score of both. The average of combined score with measurements using the Lobene Stain Index for extrinsic stain before being treated with strawberries. Hopefully the score will decreased every day starting from day 1 to day 5 of treatment.

Apigenin protects against ischemic stroke by increasing DNA repair.

Oxidative stress is an important pathological process in ischemic stroke (IS). Apigenin (APG) is a natural product with favorable antioxidative effects, and some studies have already demonstrated the antioxidative mechanism of APG in the treatment of IS. However, the mechanism of APG on DNA damage and repair after IS is not clear. The aim of this study was to investigate the mechanism of APG on DNA repair after IS. Male Sprague-Dawley rats were used to establish a model of permanent middle cerebral artery occlusion (pMCAO) on one side, and were pre-treated with gavage of APG (30, 60, or 120mg/kg) for 7 days. One day after pMCAO, the brain tissues were collected. Cerebral infarct volume, brain water content, HE staining and antioxidant index were analyzed to evaluated the brain damage. Molecular Docking, molecular dynamics (MD) simulation, immunohistochemistry, and Western blot were used to explore the potential proteins related to DNA damage repair. APG has a low binding score with DNA repair-related proteins. APG treatment has improved the volume of cerebral infarction and neurological deficits, reduced brain edema, and decreased parthanatos and apoptosis by inhibiting PARP1/AIF pathway. In addition, APG improved the antioxidative capacity through reducing reactive oxygen species and malondialdehyde, and increasing glutathione and superoxide dismutase. Also, APG has reduced DNA damage- and cell death-related proteins such as PARP1, γH2A.X, 53BP1, AIF, cleaved caspase3, Cytochrome c, and increased DNA repair by BRCA1 and RAD51 through homologous recombination repair, and reduced non-homologous end link repair by KU70. APG can improve nerve damage after IS, and these protective effects were realized by reducing oxidative stress and DNA damage, and improving DNA repair.

Amplifying protection against acute lung injury: Targeting both inflammasome and cGAS-STING pathway by Lonicerae Japonicae Flos-Forsythiae Fructus drug pair

ObjectiveAcute lung injury (ALI) is characterized by inflammation and currently lacks an efficacious pharmacological intervention. The medicine combination of Lonicerae Japonicae Flos (LJF) and Forsythiae Fructus (FF) demonstrates combined properties in its anti-infective, anti-inflammatory, and therapeutic effects, particularly in alleviating respiratory symptoms. In previous studies, Chinese medicine has shown promising efficacy in lipopolysaccharides (LPS)-induced ALI. However, there have been no reports of LJF and FF pairing for lung injury. The aim of this study is to compare the efficacy of herb pair Lonicerae Japonicae Flos-Forsythiae Fructus (LF) with LJF or FF alone in the treatment of ALI, and to explore whether LJF and FF have a combined effect in the treatment of lung injury, along with the underlying mechanism involved. MethodsA total of 36 mice were divided into six groups (control, model, LJF, FF, LF, dexamethasone) based on the treatments they received after undergoing sham-operation/LPS tracheal instillation. H&E staining and pulmonary edema indexes were used to evaluate lung injury severity. Alveolar exudate cells (AECs) were counted based on cell count in bronchoalveolar lavage fluid (BALF), and neutrophil percentage in BALF was measured using flow cytometry. Myeloperoxidase (MPO) activity in BALF was measured using enzyme-linked immunosorbent assay (ELISA), while the production of IL-1β, TNF-α, and IL-6 in the lung and secretion level of them in BALF were detected by quantitative polymerase chain reaction (qPCR) and ELISA. The effect of LJF, FF, and LF on the expression of Caspase-1 and IL-1β proteins in bone marrow derived macrophages (BMDMs) supernatant was assessed using Western blot method under various inflammasome activation conditions. In addition, the concentration of IL-1β and changes in lactatedehydrogenase (LDH) release levels in BMDMs supernatant after LJF, FF, and LF administration, respectively, were measured using ELISA. Furthermore, the effects of LJF, FF and LF on STING and IRF3 phosphorylation in BMDMs were detected by Western blot, and the mRNA changes of IFN-β, TNF-α, IL-6 and CXCL10 in BMDMs were detected by qPCR. ResultsLF significantly attenuated the damage to alveolar structures, pulmonary hemorrhage, and infiltration of inflammatory cells induced by LPS. This was evidenced by a decrease in lung index score and wet/dry weight ratio. Treatment with LF significantly reduced the total number of neutrophil infiltration by 75% as well as MPO activity by 88%. The efficacy of LF in reducing inflammatory factors IL-1β, TNF-α, and IL-6 in the lungs surpasses that of LJF or FF, approaching the effectiveness of dexamethasone. In BMDMs, the co-administration of 0.2 mg/mL of LJF and FF demonstrated superior inhibitory effects on the expression of nigericin-stimulated Caspase-1 and IL-1β, as well as the release levels of LDH, compared to individual treatments. Similarly, the combination of 0.5 mg/mL LJF and FF could better inhibit the phosphorylation levels of STING and IRF3 and the production of IFN-β, TNF-α, IL-6, and CXCL10 in response to ISD stimulation. ConclusionThe combination of LJF and FF increases the therapeutic effect on LPS-induced ALI, which may be mechanistically related to the combined effect inhibition of cyclic-GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) and NOD-like receptor family protein 3 (NLRP3) inflammasomes pathways by LJF and FF. Our study provides new medicine candidates for the clinical treatment of ALI.

Lipopolysaccharide modification enhances the inhibitory effect of clodronate liposomes on hepatic fibrosis by depletion of macrophages and hepatic stellate cells

Hepatic fibrosis is a complex chronic liver disease in which both macrophages and hepatic stellate cells (HSCs) play important roles. Many studies have shown that clodronate liposomes (CLD-lipos) effectively deplete macrophages. However, no liposomes have been developed that target both HSCs and macrophages. This study aimed to evaluate the therapeutic efficacy of lipopolysaccharide-coupled clodronate liposomes (LPS-CLD-lipos) and the effects of liposomes size on hepatic fibrosis. Three rat models of hepatic fibrosis were established in vivo; diethylnitrosamine (DEN), bile duct ligation (BDL), and carbon tetrachloride (CCl4). Hematoxylin and eosin staining and serological liver function indices were used to analyze pathological liver damage. Masson's trichrome and Sirius red staining were used to evaluate the effect of liposomes on liver collagen fibers. The hydroxyproline content in liver tissues was determined. In vitro cell counting kit-8 (CCK-8) and immunofluorescence assays were used to further explore the effects of LPS modification and liposomes size on the killing of macrophages and HSCs. Both in vitro and in vivo experiments showed that 200 nm LPS-CLD-lipos significantly inhibited hepatic fibrosis and the abnormal deposition of collagen fibers in the liver and improved the related indicators of liver function. Further results showed that 200 nm LPS-CLD-lipos increased the clearance of macrophages and induced apoptosis of hepatic stellate cells, significantly. The present study demonstrated that 200 nm LPS-CLD-lipos could significantly inhibit hepatic fibrosis and improve liver function-related indices and this study may provide novel ideas and directions for hepatic fibrosis treatment.

Oncostatin M [OSM] protein immuno-expression in oral squamous cell carcinoma – A prospective study

BackgroundAn important factor in the progression of tumours is the formation of blood vessels in the tumour stroma. When there is an upregulation of angiogenesis stimulators or a downregulation of angiogenesis inhibitors, the tumour may manifest its capacity for angiogenesis. Immune cells secrete a substance called Oncostatin M (OSM), which has been linked to cancer angiogenesis and subsequent pathogenesis. This study is aimed to assess OSM expression in oral squamous cell carcinoma. ObjectiveTo compare the intensity of expression of Oncostatin M [OSM] in normal oral mucosa and Oral Squamous Cell Carcinoma and to analyse their role in the disease progression by enhancing angiogenesis and metastasis. Materials and methodsThe study analysed the expression of OSM with the immunohistochemical aid. The samples were treated with Anti-OSM polyclonal antibody and immunohistochemical staining was done using NovoLinkTM NovocastraTM Polymer Detection System (Leica Microsystems). Stained samples were categorized based on the staining percentage and staining intensity. ResultsThe comparison of staining percentage between the epithelium of control group and the OSCC group revealed that it was statistically significant [p value = 0.002685]. Similarly, the comparison of staining intensity [p value = 0.001202] and staining index [p value = 0.001817] between the control group and the OSCC group revealed that it was statistically significant. ConclusionThe expression of oncostatin M in normal epithelium and OSCC group with the given immunohistochemical staining percentage, intensity and staining index suggests strong diagnostic potential of oncostatin M which can also be used for further therapeutic.

HAPLN1 knockdown inhibits heart failure development via activating the PKA signaling pathway

BackgroundHeart failure (HF) is a heterogeneous syndrome that affects millions worldwide, resulting in substantial health and economic burdens. However, the molecular mechanism of HF pathogenesis remains unclear.MethodsHF-related key genes were screened by a bioinformatics approach.The impacts of HAPLN1 knockdown on Angiotensin II (Ang II)-induced AC16 cells were assessed through a series of cell function experiments. Enzyme-linked immunosorbent assay (ELISA) was used to measure levels of oxidative stress and apoptosis-related factors. The HF rat model was induced by subcutaneous injection isoprenaline and histopathologic changes in the cardiac tissue were assessed by hematoxylin and eosin (HE) staining and echocardiographic index. Downstream pathways regulated by HAPLN1 was predicted through bioinformatics and then confirmed in vivo and in vitro by western blot.ResultsSix hub genes were screened, of which HAPLN1, FMOD, NPPB, NPPA, and COMP were overexpressed, whereas NPPC was downregulated in HF. Further research found that silencing HAPLN1 promoted cell viability and reduced apoptosis in Ang II-induced AC16 cells. HAPLN1 knockdown promoted left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS), while decreasing left ventricular end-systolic volume (LVESV) in the HF rat model. HAPLN1 knockdown promoted the levels of GSH and suppressed the levels of MDA, LDH, TNF-α, and IL-6. Mechanistically, silencing HAPLN1 activated the PKA pathway, which were confirmed both in vivo and in vitro.ConclusionHAPLN1 knockdown inhibited the progression of HF by activating the PKA pathway, which may provide novel perspectives on the management of HF.