Staining Index Research Articles

Effect of sodium hyaluronate eye drops combined with tobramycin, dexamethasone and pranoprofen eye drops in the treatment of dry eye after phacoemulsification.

To investigate the clinical effect of sodium hyaluronate eye drops combined with tobramycin, dexamethasone, and pranoprofen eye drops in the treatment of dry eye after phacoemulsification. Medical records of 188 patients with dry eye after phacoemulsification, treated in our hospital from August 2020 to August 2021, were included and divided into groups based on the type of dry eye treatment. Patients in the control group (n = 90) were treated with tobramycin, dexamethasone, and pranoprofen eye drops. Patients in the test group (n = 98) were treated with sodium hyaluronate, tobramycin, dexamethasone, and pranoprofen eye drops. The tear secretion test (TST), film rupture time (BUT), corneal fluorescence staining (FS) score, levels of interleukin (IL)-6, and tumor necrosis factor (TNF)-α before the treatment and at 1 month follow-up, the overall total effective rate and the number of completely cured cases were compared between the two groups. Multiple linear regression was used to calculate coefficients for predicting clinical variables. Ordinal logistic regression was used to compute coefficients and odds ratios for predicting effective scores. The total effective rate in the observation group (99%) was significantly higher than that in the control group (80.00%). After the treatment, the TST and BUT indexes of the observation group were higher compared to the control group, and the FS score of the observation group was lower than that of the control group. TNF-α and IL-6 levels of the observation group were lower compared to the control group. Logistic regression analysis demonstrated that the differences in the effective rate and clinical variables between the two groups remained significant after adjusting for the effect of age. A combination of sodium hyaluronate, tobramycin, dexamethasone, and pranoprofen eye drops for the treatment of dry eye after phacoemulsification can improve the curative effect, reduce corneal fluorescein staining and inflammatory factor index levels compared to the treatment regimen lacking sodium hyaluronate eye drops.

Qingdu decoction can reduce LPS induced ACLF endotoxemia by regulating microRNA-34c/MAZ/TJs and microRNA-122a/Zonulin/EGFR signal pathway

Ethnopharmacological relevanceQingdu Decoction (QDT) is a traditional Chinese medicine (TCM) that was derived from Xiaochengqi Decoction, a famous decoction documented in the book of Treatise on Exogenous Febrile Disease in the Eastern Han Dynasty. According to our years of clinical application, QDT showed satisfactory efficacy in the treatment of endotoxemia in acute-on-chronic liver failure (ACLF). However, the underlying molecular mechanisms remain largely unknown. Aim of studyIn this study, we aimed to systematically evaluate the intervention effect of QDT on endotoxemia in rats and further clarify its potential regulatory mechanism. Materials and methodsThe rat model of ACLF endotoxemia was induced by TAA and LPS + D-Gal. Then the rats were treated with clinical doses of QDT and lactulose. The rats were divided into four groups: CG, MG, QG and LG. The target microRNA was screened by high-throughput sequencing. The rat weight, liver index, hepatointestinal phenotype, serum biochemical indexes, mast cell activity, and hepatointestinal histopathology were used to evaluate the intervention effect. Western blot analysis was used to detect the expression levels of MAZ and its downstream genes ZO-1 and Occludin, and the expression levels of Zonulin and its downstream gene EGFR in colon. Finally, the expression of the miR-34c, MAZ, ZO-1, Occludin, miR-122a, Zonulin, and EGFR in colon was detected by qRT-PCR to further confirm the mechanism of the miR-34c/MAZ/TJs pathway and the miR-122a/Zonulin/EGFR pathway. ResultsThe rat weight, liver index, liver and colon phenotype, and serum biochemical indexes showed that QDT could significantly reduce liver and intestine injury and inhibit the progress of ACLF and endotoxemia. Toluidine blue staining and cytokine indexes showed that QDT could inhibit the activity of MCs and reduce the release of inflammatory factors. Mechanistically, QDT can inhibit the activity of MCs, activate miR-34c/MAZ/TJs pathway and miR-122a/Zonulin/EGFR pathway in colon, promote the recovery of intestinal barrier homeostasis, reduce and restore the damage of endotoxemia. ConclusionOur results suggested that QDT can significantly reduce rat ACLF endotoxemia by regulating the miR-34c/MAZ/TJs pathway and the miR-122a/Zonulin/EGFR pathway in colon.

AML MRD By Multiparameter Flow Cytometry Using Laip/Dfn and LSC: Methodological Aspects in a Multicentric Study of the French-Flow MRD AML ALFA Network

Introduction: MRD follow-up is now mandatory for treatment response evaluation in AML clinical trials (ELN 2017 and 2021 guidelines: Heuser et al,Blood doi:10.1182/blood.2021013626.). Beside to molecular biology methods, multiparameter flow cytometry assay represents the most reliable approach. The aim of this study was to implement an harmonized follow up of the patients using a standardized MRD flow approach across 30 French hematology laboratories participating in AML clinical trials. Methods: To obtain superposable results, the network has established recommendations from Wet labs procedure to clinical reports using common panels, flow cytometers settings and analysis strategy. We designed a 2-tubes panel with minimal mandatory 8c markers by tube, according to ELN recommendations to identify the pattern of LAIP/DfN in bulk cells and the LSC (Leukemia Stem Cells) in CD34+CD38- fraction. A « backbone » CD34/CD38/CD45/CD117 was used, completed by CD7,CD56, CD13, CD33, CD19 for the 1st tube and CD90(Thy-1), Mix (CD97+CLL1+TIM3), CD123, CD45RA for the 2nd one. This panel could be used in 8c,10c,12c (HLADR, CD36, CD10, CD200) implemented on Becton Dickinson (CANTO and LYRIC) or Beckman Coulter (NAVIOS and DxFlex) cytometers (Fig 1A). Harmonization of the sensitivity between platforms (18 BD and 12 BC) was performed using 8 Picks Rainbows calibration beads. Immunostaining was performed after "bulk" lysis technique (NH4Cl). A total of minimal 500 000 cells was acquired in each tube and data were analyzed using DIVA/FACSUITE and KALUZA software. Results: To detect any bias between the platforms, firstly we tested 10 EDTA fresh regenerative marrow samples in parallel at 2 platforms (CANTO and NAVIOS). Similarity of the staining index between positive and negative population were compared between the samples and the 2 platforms showing no differences. Secondly, regular QC (Quality Control Sample), of normal bone marrow (from healthy donor) 1/year (a total of 5 since 2017) was shared among the participants centers(Fig1B).The specific events populations (nHSC, MPP, LMPP"like,etc) were evaluated by centers using a common gating strategy. In a third step, standardization of the data analysis strategy obtained in the centers was subsequently evaluated using a virtual quality control(CQA) by sharing MRD FCS data files. About 90% of the locally analyzed data were included in an interval centralized on the average of the series mean +/- 2 SD. During the follow up of the patients, a series of 18 Web educational meeting was implemented to review and validate cases with difficulties regarding analysis interpretation and confrontation with molecular MRD (transcripts, NPM1, WT1, NGS) and also chimerism in allograft follow-up (mainly cases with clonal hematopoiesis or stressed bone marrow). Finally, the harmonized data generated in our group allowed to perform a Computer aided design (CAD) in assessment of AML MRD flow using proprietary developed R script based upon FlowSom. It performs an automated definition of metacluster with abnormal population (by comparison to a set of reference bone marrow) which are quantified in the CD34+ CD117+ space for all samples (diagnosis, follow up and references marrow). It then extracts the metacluster significantly different from that observed for the reference samples (> mean+6sd. It calculates the MRD in % of WBC CD45+. The limits of sensitivity of the CAD method shows that a MRD remains evaluable by CAD up to 0.04% (50 events in 1 000 000). We compared the results between the conventional and the CAD method on 60 MRD samples. The slope of the correlation line was 0.95 with an R2 of 0.97 (Fig 1C). Using the definition threshold of a positive MRD at 0.1%, the agreement of the results between the two methods was evaluated by the Cohen kappa coefficient of 0.87. Conclusions: This methodological validation protocol is a mandatory step to consider the use of MRD flow in AML clinical trials. Choosing a multicentric approach could be challenging, but our first results are promising and showed the feasibility of this concept when: (i) a straight harmonisation of the instruments sensitivity and samples preparation are established, and (ii) training and systematic education among the analytical operators are regularly performed. Finally, our pilot study shows a very strong correlation between conventional method and unsupervised analyses using data generated by the multicentric network. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Detection, Monitoring and Cell Kinetics of Circulating CAR-T Cells in B-Cell Malignancies

Introduction CAR-T cell therapy has proven to be a disruptive treatment in the hematology field. Tisagenlecleucel and Axicabtagene ciloleucel showed impressive efficacy in relapsed/refractory B-cell lymphoma; however, less than 50% of patients maintain a long-term response and predictors of outcome are still inconsistently defined. Tisa-cel and Axi-cel kinetics have been poorly investigated in real life settings so it becomes necessary to evaluate possible biomarkers that predict response or toxicity. In clinical trials, the efficacy of the treatment is determined by the in vivo expansion and persistence of CAR-T cells. Real life evaluations assessing the immunophenotypic composition of CAR-T cell infusion products and analyzing how these specific features affect CAR-T cell expansion and persistence in vivo and thus outcome, are scarce. Here, we aimed to optimize the detection of CD19 CAR-T cells in blood and to identify phenotypic features associated with post-infusion expansion and outcomes. Methods CD19 CAR-T cells were monitored by flow cytometry from peripheral blood in 48 patients with diffuse large B-cell lymphoma, transformed follicular lymphoma, primary mediastinal B cell lymphoma or B-cell acute lymphoblastic leukemia. Flow cytometry approach was performed for determining cell kinetics (n=48) at days 5, 7, 11, 14, 17 and 20 (±1) after CAR-T cells infusion. In parallel, immunophenotype characterization of CAR- T lymphocytes was conducted at the day of CAR-T expansion in blood. Results After testing different commercially available antibodies for the detection of CD19 CAR-T cells, CD19 protein conjugated with biotin and identified by streptavidin showed the highest resolution in contrast to Antigen CD19 labeled FITC (stain index of 22.19 vs 4.64, p=0.0286). The median day of peak expansion of CAR-T cells was 7 days (range of 5-14 days) after infusion, reaching a median of 119 CAR+ cells/uL (range of 2.4.-1019 CAR+ cells/uL) in blood. Analysis of immune-phenotypic profile of overall circulating T lymphocytes displayed no significant differences in the distribution of naive/memory/effector from CD4+ and/or CD8+ subpopulations between non-CAR T (normal T lymphocytes) and CAR-T cells. However, CAR-T cells showed a significantly higher expression levels of activation (CD69+), proliferation (Ki67+), cytotoxicity (CD107a+) and exhaustion (PD1+TIM3+, PD1+LAG3+) markers in comparison to T lymphocytes (p<0.05). Overall, 31.6% (6/19) patients reached complete remission while 47.4% (9/19) achieved partial response at the first month after CAR-T therapy with event free survival (EFS) of 73% at 1 year (median follow-up of 1 year). Remarkably, cases with lower expression levels of PD-1 and LAG-3 (i.e <5.2%) in CD4+CAR+ T cells at the day of peak expansion had a worse outcome (EFS at 1 year 44% vs 90% for patients with low vs high expression of the above exhaustion markers, p=0.04) (Figure 1A). Similarly, patients with a highly cytotoxicity pattern (i.e CD107a expression >6%) within CD8+CAR+ T cells had a shorter EFS (50%) as compared to those with <6% of CD8+CAR+CD107a+ (89%) (p=0.06) (Figure 1B). Finally, patients with >3.2% CD4-CD8- T cells within the nonCAR T lymphocytes showed superior outcome as compared to those patients who displayed lower levels of double-negative T cells (100% vs 50% of EFS rates, respectively) (p=0.03). Conclusions In our cohort, the median day of CD19 CAR-T cell peak expansion as assessed by flow cytometry was 7 days after infusion. Markers of exhaustion within the CAR+ population and percentage of double-negative T lymphocytes allows an early identification of prognostic groups in order to guide therapeutic actions. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Association between poor sleep quality and an increased risk of dry eye disease in patients with obstructive sleep apnea syndrome.

Obstructive sleep apnea (OSA) is related to an increased incidence of dry eye disease (DED). However, their exact relationship is unknown and requires further well-designed studies with advanced mechanisms detection. This case-control study included 125 OSA cases and 125 age-gender-matched controls enrolled in the hospital between 1 January and 1 October 2021. OSA diagnosis and classification were performed using a polysomnography (PSG) assay. Detailed ophthalmological examinations, including the Schirmer I test, corneal staining, and ocular surface disease index (OSDI), were used to detect DED-related parameters. A comprehensive ocular surface assay was performed to measure a series of parameters, including first non-invasive first tear film break-up time (f-NIBUT), average non-invasive first tear film break-up time (av-NIBUT), tear meniscus height (TMH), and loss of meibomian gland. In addition, the Pittsburgh Sleep Quality Index (PSQI) scale was used to assess sleep quality. Compared to the control, the OSA group showed an increased DED risk (P = 0.016) along with an increased PSQI score and a higher rate of poor quality sleep (P < 0.001 and P = 0.007, respectively). Stratification of OSA cases indicated that DED-related parameters were impaired in patients with severe OSA (P < 0.05). The analysis of DED-parameters-related factors showed significant correlations between OSA-related indexes and PSQI (P < 0.05). Moreover, the poor sleep quality group in the OSA cases showed worse DED-related parameters (P < 0.05), which was not observed in the control group. OSA, especially the severe stage OSA, was related to an increased risk of DED. Also, sleep quality was correlated with the onset of both OSA and DED, where poor sleep quality revealed a relationship between OSA and the risk of DED. Overall, our findings provided evidence for advanced management of DED and OSA in future.

Relationship between Black Chromogenic Stains and Hemoglobin Levels

Background: Occurance of black discolouration of teeth is very common and a clinical challenge in the present day dentistry. Stains associated with chromogenic bacteria have a high rate of recurrence even after thorough scaling and polishing procedures. The mechanism of stain formation is still not understood clearly though its chemical composition is mainly iron suiphide and bacteria like actinomyces have been identified in plaque samples of these patients which have the ability to form iron sulphides. The iron content of saliva is known to increase in iron deficiency anemia and iron overload. Iron deficiency anemia is also associated with low hemglobin levels. Aim: To determine the hemoglobins levels in patients with and without black chromogenic stains. Methodology: A total of 40 patients of age group 15-40 years were selected and divided in to two groups. Group ‘A’ (Test group) consisted of 20 patients with black stains on atleast 8 tooth. Group “B” (Control group) consisted of 20 patients without any stains on the tooth surfaces. The individuals were screened for Lobenes stain index, Oral hygiene index and haemoglobin levels. The scores were analysed using chi –square tests. Results: It was observed that there was no significant difference between the haemoglobin levels of patients with and without stains. The oral hygiene scores were found to be significantly higher in subjects with stains compared to controls. Conclusion: The study suggests that the presence of stains were not associated with the reduced haemoglobin levels. The chromogenic black stains were more prevalent among subjects with good oral hygiene.

2'-Fucosyllactose Suppresses Angiogenesis and Alleviates Toxic Effects of 5-Fu in a HCT116 Colon Tumor-Bearing Model.

The present study was aimed at examining the anti-tumor effects and molecular mechanisms of 2'-fucosyllactose (2'-FL). At the beginning, the viabilities of four types of colon cancer cells were analyzed after exposure to increasing concentrations of 2'-FL, and HCT116 cells were selected as the sensitive ones, which were applied in the further experiments; then, interestingly, 2'-FL (102.35 µM) was found to induce apoptosis of HCT116 cells, which coincides with significant changes in VEGFA/VEGFR2/p-PI3K/p-Akt/cleaved Caspase3 proteins. Next, in a tumor-bearing nude mouse model, HCT116 was chosen as the sensitive cell line, and 5-fluorouracil (5-Fu) was chosen as the positive medicine. It was noteworthy that both 2'-FL group (2.41 ± 0.57 g) and 2'FL/5-Fu group (1.22 ± 0.35 g) had a significantly lower tumor weight compared with the control (3.87 ± 0.79 g), suggesting 2'-FL could inhibit colon cancer. Since 2'-FL reduced the number of new blood vessels and the malignancy of tumors, we confirmed that 2'-FL effectively inhibited HCT116 tumors, and its mechanism was achieved by regulating the VEGFA/VEGFR2/PI3K/Akt/Caspase3 pathway. Moreover, though HE staining and organ index measurement, 2'-FL was validated to alleviate toxic effects on liver and kidney tissue when combining with 5-Fu. In conclusion, 2'-FL had certain anti-tumor and detoxification effects.

Erythrocyte lysing solutions have a detrimental effect in flow cytometric dendritic cell detection.

Flow cytometry (FCM) enumeration of peripheral blood dendritic cells (PBDCs) is a minimally invasive procedure extremely useful for immunological studies. Numbers of PBDCs vary depending on age, lifestyle, or in pathologies like cancer, leukemia or immunodeficiencies. Conventional methods for PBDC identification by FCM involve red blood cell lysis using either formaldehyde or ammonium chloride-based solutions. This specific procedure has been widely reported to cause a detrimental effect as well as an artifactual detection of target populations. Alternatively, minimal sample perturbation assays that avoid the use of erythrolytic solutions with centrifugation steps and preserve the native cellular state are simpler and more robust than conventional methods. In this study, we aimed to evaluate how conventional FCM assays can alter dendritic cell (DC) counting when compared with minimal sample perturbation protocols, in terms of absolute cell counting, percentage and stain index (SI) of PBDC subsets. We evaluated the use of three different erythrolytic solutions (CyLyse, OptiLyse C, and Pharm Lyse) on a series of n= 20 peripheral blood specimens for conventional and plasmacytoid DCs detection as well as for leukocyte and basophil detection. Our results showed a significant reduction of leukocytes and specifically, of DCs and basophils in terms of absolute number when using erythrolytic solutions. In conclusion, our study shows that PBDC counting is heavily affected when lysing solutions are used, indicating that these stellate-shaped populations appear to be more labile.

Effect on Dental Stains by Potassium Tripolyphosphate Added Chewing Gum

Background: Today, people worldwide consider the discoloration of teeth the main concern, therefore, dental stains are an important problem for a lot of patients, especially for smokers, and tea and coffee consumers. Objective: This trial was planned to evaluate the effectiveness of a sugar-free chewing gum added with potassium tripolyphosphate, compared to a placebo chewing gum on the development or the removal of dental extrinsic stains preserving regular daily oral hygiene. Methods: This was a single-center, double-blind, randomized, 6-week parallel controlled clinical trial. Among those who were eligible for the trial, 162 adult participants were randomly allocated into two groups of 81 each and were instructed to maintain customary oral hygiene. All subjects started the trial period after an in-office dental visit to set the stain index baseline. They chewed one of the two chewing gums for six weeks, five pieces per day, preferably after meals and snacks, for 10 minutes. Both chewing gums were sugar-free, 2g of weight with the same size and shape. The test chewing gum contained potassium tripolyphosphate (24.4 mg per piece), the control chewing gum was identical without potassium tripolyphosphate, therefore, it did not contain any anti-stain agent. The dental extrinsic stain was measured at the first visit and at the end of six weeks by the Modified Lobene Stain Index (MLSI). Comparisons between the groups were performed using ANOVA after adjustment of the baselines, and comparisons between initial and final indexes inside the groups were performed using paired t-tests. Results: After the 6 weeks, 154 subjects completed the trial, 77 in each group. The mean difference in stain composite index for all sites after six weeks was 0.04±0.07 in the control group and -0.03±0.07 in the test group. This difference was statistically significant after baseline adjustment (p&lt;0.001). Moreover, the differences in stain indexes for both buccal or lingual-palatal sites showed a statistically significant difference (p&lt;0.001) for those using the test chewing gum versus the control chewing gum. Conclusion: The overall findings of this clinical study suggest that the use of chewing gum containing potassium tripolyphosphate can reduce dental stains versus placebo chewing gum on frontal teeth after six weeks of maintaining regular oral hygiene with normal tooth brushing.

P017 Echinocandin resistance mechanism in Candida tropicalis and Candida glabrata

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PMObjective Candida tropicalis and Candida glabrata account for 41.6% and 7.08% of total Candidaemia cases in India. Echinocandins are the first-line treatment option for these infections. Resistance to Echinocandins is rare with Candida sp. However, in recent years, has been noted across many centers. We determined the mechanism of echinocandin resistance in C. tropicalis and C. glabrata.Methods C. tropicalis and C. glabrata isolated from Candidaemia patients over a period of 3 years (August 2016-July 2019), identified by MALDI-TOF-MS were used in this study. Antifungal susceptibility testing was done following CLSI broth microdilution reference method (M 27A). FKS1 gene was sequenced using species-specific primers for the presence of any mutation. To determine any changes in the cell wall chitin and glucan contents, expression fold changes of chitin synthase (CHS1, CHS2, and CHS3), and glucan synthase genes upon caspofungin treatment were determined using real-time qPCR. These findings were correlated with cell wall chitin and glucan content determined by flowcytometry.ResultsA total of 3558 Candida species were isolated from patients of all age groups at our hospital. C. tropicalis was the predominant agent (34%), while the prevalence of C. glabrata was 6%. A total of 17 (8.5%) C. glabrata and 3 (0.25%) C. tropicalis exhibited reduced susceptibility to echinocandins. All these isolates carried a wild-type FKS genes.In C. tropicalis, inducible expression of Chs1, Chs2 and Chs3 genes were comparable among susceptible and resistant isolates1.8 (0.4-2.8) vs. 2.5 (0.9-6.6), P = .247); [0.7 (0.3-1.8) vs. 0.7 (0.2-1.6), P = .793]; [1.3 (0.14-4.8) vs. 1.1(0.48-1.7), P = .522], respectively. In concordance with gene expression, there was no significant difference in cell wall chitin contents among resistant and susceptible [14.37 (6.5-24.8) vs 16.28 (6.0-24.7), P = .114] C. tropicalis isolates. In contrast in resistant isolates of C. glabrata, caspofungin treatment resulted in significantly higher induction of chitin synthase genes compared to susceptible isolates; Chs1 [2.34 (0.24-9.71) vs. 1.56 (0.55-4.5) (P = .007)], CHS2 [1.59 (0.33-8.0) vs. (2.3 (0.69-6.15), P = .0006], and CHS3 gene [3.8 (0.13-12.73) vs. 1.9 (0.56-7.16), P <.0001]. Flowcytometric data in terms of chitin content, correlated well with expression changes as staining index was significantly higher in resistant compared to susceptible isolates [320 (198-535) vs. 164 (5.34-254.10) P = .0001]. Glucan synthase expression was comparable in susceptible and resistant isolates of C. tropicalis [3.47 (1.57-7.63) vs. 4.41 (0.41-17.51), (P = .518)]. However, glucan synthase gene expression was found significantly increased in resistant C. glabrata isolates compared to susceptible isolates; 3.10 (1.02-16.45) vs. 1.61 (0.13-7.67), P <.001.ConclusionWe evaluated the role of cell wall components in echinocandin resistance in isolates with reduced susceptibility to echinocandins but lacking an FKS1 mutation. While chitin was induced at higher levels in C. glabrata, a similar finding was not observed in C. tropicalis. This warrants further studies to elucidate the role of fungal cell wall polymers in resistance.

IMPACT OF BAKING SODA ON TEETH WHITENING AMONG TOBACCO USERS - AN INTERVENTIONAL STUDY

Background: A more stylish and wonderful grin has been a typical longing for a great many people looking for dental treatment. Tooth tint is considered as a fundamental factor in the dental charisma, especially in the front upper dentition. Tooth staining might be brought about by inherent or outward stain or a blend of tooth. Baking soda is the successful and simple strategy to improve brightening of teeth. As a decisive assurance of the nurse, it is one of responsibility to dealing with the patient's consideration to survey the oral mucosa and settle on resulting techniques for oral cleanliness and improving teeth brightening. Methods and methodology: The present study aims to assess the effectiveness of baking soda on whitening of teeth among tobacco users. A quantitative quasi experimental research design was conducted among 60 tobacco users. Purposive sampling technique was used to select samples. Semi-structured interview was used to collect demographic data and teeth whitening was assessed using Modified Lobene Stain Index. The tobacco users in experimental group were given baking soda to brush the teeth two minutes per day. After one week, the tooth was re-assessed. Results: The study results show significant improvement in the teeth whitening among experimental group than control group after the intervention at the level of p&lt;0.001. Conclusion: This reveals that baking soda is highly significant in the experimental group because it improves teeth whitening of the tobacco users. This indicates that baking soda is the effective and easy method to improve teeth whitening.

Magnetic resonance spectroscopy associations with clinicopathologic features of estrogen-dependent endometrial cancer

BackgroundWe studied the magnetic resonance spectroscopy (MRS) associations with clinicopathologic features of estrogen-dependent endometrial cancer (type I EC).MethodsTotally 45 patients with type I EC who underwent preoperative multi-voxel MRS at 3.0 T were enrolled. The mean ratio of the Cho peak integral to the unsuppressed water peak integral (Cho/water) of the tumor was calculated. The Cho/water and apparent diffusion coefficient (ADC) of type I EC with and without local invasion, as well as with different levels of Ki-67 staining index (SI) (≤ 40% and > 40%), were compared. Correlation test was used to examine the relationship of Cho/water, as well as mean ADC, with Ki-67 SI, tumor stage, and tumor grade.ResultsThe mean Cho/water of EC with Ki-67 SI ≤ 40% (2.28 ± 0.93) × 10−3 was lower than that with Ki-67 SI > 40% (4.08 ± 1.00) × 10−3 (P < 0.001). The mean Cho/water of EC with deep and superficial myometrial invasion was (3.41 ± 1.26) × 10−3 and (2.43 ± 1.11) × 10−3, respectively (P = 0.011). There was no significant difference in Cho/water between type I EC with and without cervical invasioin ([2.68 ± 1.00] × 10−3 and [2.77 ± 1.28] × 10−3, P = 0.866). The mean Cho/water of type I EC with and without lymph node metastasis was (4.02 ± 1.90) × 10−3 and (2.60 ± 1.06) × 10−3, respectively (P = 0.014). The Cho/water was positively correlated with the Ki-67 SI (r = 0.701, P < 0.001). There were no significant differences in ADC among groups (all P > 0.05).ConclusionMRS is helpful for preoperative assessment of clinicopathological features of type I EC.

EFEKTIVITAS PASTA GIGI BAKING SODA DAN NON BAKING SODA TERHADAP PENURUNAN STAIN PADA PEROKOK

Many people do not realize that smoking habits can affect the level of dental and oral hygiene, smoking causes abnormalities of the tongue, gums, oral mucosa, palate in the form of nicotinic stomatitis, fungal infections, and causes discoloration of the te eth. Sodium bicarbonate is a chemical compound with the formula NaHCO3 belonging to the salt group and is often found in the form of white powder crystals. Where the content of sodium bicarbonate can remove stains on smokers. The purpose of this study was to determine the effectiveness of the use of toothpaste containing baking soda and non-baking soda to reduce stains in smokers.This type of research is analytic with a quasi-experimental method. The sample was used as many as 62 people then divided into 2 groups which were then checked for staining conditions before and after the use of toothpaste containing baking soda and non -baking soda.The results showed that the average tooth staining before and after brushing using toothpaste containing baking soda showed a decrease in staining on the Lobene intensity measurement as much as 0.324, lobene area as much as 0.217, combined as much as 0.541, with p = 0.000 &lt; = 0, 1. While the average tooth staining before and after using non -baking soda toothpaste there was a decrease in stain on Lobene intensity measurements as much as 0.043, Lobene area as much as 0.016, combined as much as 0.059 , with p value = 0.116 &gt; = 0.1.This study concluded that the use of toothpaste containing baking soda was more effective than non-baking soda toothpaste to reduce the stain index in smokers. It is suggested that the public can use toothpaste containing baking soda as an alternative to remove stains on the teeth, and pay more attention to the status of oral hygiene.

Development of a 27-color panel for the detection of measurable residual disease in patients diagnosed with acute myeloid leukemia.

Acute myeloid leukemia (AML) measurable residual disease (MRD) evaluated by multiparametric flow cytometry (MFC) is a surrogate for progression-free and overall survival in clinical trials and patient management. Due to the limited number of detection channels available in conventional flow cytometers, panels used for assessing AML MRD are typically split into multiple tubes. This cripples the simultaneous and correlated assessment of all myeloblast measurements. In response, we prototyped a single-tube 27-color MFC assay for the evaluation of AML MRD, incorporating all recommended markers. Marrow aspirates from 22 patients were processed for analysis using full spectrum flow cytometry (FSFC). The signal resolution of each marker was compared between samples stained with single antibody vs. the fully stained panel. The analytical accuracy for quantifying hematopoietic cells between our established 8-color assay and the new 27-color method were compared. Variations within an operator and between separate operators were assessed to evaluate the assays reproducibility. The limited of blank (LOB), limit of detection (LOD), and lower limit of quantification (LLOQ) of the 27-color method were empirically determined using limiting dilution experiments. The stability of antibody cocktails over a period of 120 h was also studied using cryopreserved marrow cells. The stain indices for all antibodies were lower in the fully stained panel compared to cells stained with one antibody but clear separations between negative and positive signals were achieved for all antibodies. Our results demonstrated a high concordance between the established 8-color method and the new 27-color assay for enumerating myeloblasts and MRD interpretation within and between operators. The data further showed that the single-tube 27-color assay easily achieved the minimum required detection sensitivity of 0.1%. When antibodies were combined, however, expression intensity of some antigens deteriorated significantly when stored. Our single-tube 27-color panel is a suitable, high sensitivity flow cytometric approach that can be used for AML MRD testing, which improves the correlation of aberrant antigens and detection of asynchronous differentiation patterns. Based on the stability study, we recommend the full panel be made prior to staining.

Expression of Breast Cancer Gene 2 in Oral Squamous Cell Carcinoma and its Correlation with the Metastatic Potential

Background: Breast cancer gene 2 (BRCA2), a tumor suppressor gene, has been extensively evaluated in various cancers and correlated with metastatic potentials. However, to the best of our knowledge, the correlation has not been reflected in the literature in patients with oral squamous cell carcinoma (OSCC). Any positive correlation may yield the protein as an exploitable marker for the prediction of metastasis and target for chemotherapy in patients with OSCC. Materials and Methods: This observational retrospective study was conducted on 54 tissue samples, 18 samples each of normal oral mucosa, and of OSCC with and without cervical lymph node metastasis. The study samples were immunohistochemically stained with BRCA2 antibody. The staining index was calculated as a product of the percentage of positive cells per high-power field (HPF) (A) and the staining intensity (B). Results: Based on intergroup comparisons, it was found that the percentage of positive cells/HPF, staining intensity, and staining index were higher in OSCCs with cervical lymph node metastasis and lower in normal oral mucosa samples. It was found that a large number of the normal oral mucosa samples (n = 13) did not show any BRCA2 immunoreactivity and a large number of the OSCC samples with cervical lymph node metastasis displayed high BRCA2 immunoreactivity (n = 15). Conclusion: Overexpression of BRCA2 is linked to the presence of metastases in patients with OSCC and has the potential to be utilised as a marker to predict the outcome of the patient's treatment. The findings may be used in the future to develop targeted therapies, depending on the direction that future research follows.

FOSL1 knockdown ameliorates DSS-induced inflammation and barrier damage in ulcerative colitis via MMP13 downregulation.

Ulcerative colitis (UC) is a chronic inflammatory disease of the colon. Fos-related antigen 1 is highly expressed in mild UC and affects the maintenance and delayed recurrence of inflammatory bowel disease. Therefore the present study aimed to investigate the role of Fos-like antigen-1 (FOSL1) in UC. Use of the JASPAR database predicted the possible interaction of FOSL1 and the MMP13 promoter. FOSL1 and MMP13 mRNA and protein expression levels in dextran sodium sulfate (DSS)-induced HT29 cells and colon tissues of a UC mice model were determined using reverse transcription-quantitative PCR and western blotting, respectively. MMP13 promoter activity was determined using the dual-luciferase reporter assay, the relationship between FOSL1 and MMP13 promoter was determined using chromatin immunoprecipitation, inflammatory factor levels were quantified using ELISA, cell monolayer permeability analysis was performed using transepithelial electrical resistance measurements and tight junction protein expression levels were determined by western blotting. After constructing a UC mice model, mice colonic injury was determined with the quantification of colon length, H&E staining and disease activity index. The results demonstrated that FOSL1 and MMP13 were upregulated in DSS-induced HT29 cells and tissues. FOSL1 was also determined to be bound to the MMP13 promoter region and was demonstrated to upregulate MMP13 expression levels. Furthermore, FOSL1 knockdown inhibited DSS-induced inflammation and barrier damage in HT29 cells via MMP13 downregulation. FOSL1 knockdown also ameliorated colonic injury, inflammation and barrier damage in the tissues of the UC mice model via MMP13 downregulation. Overall, FOSL1 knockdown was demonstrated to potentially ameliorate DSS-induced inflammatory injury and protect the intestinal barrier integrity in the UC mice model via MMP13 downregulation.

Evaluation of Expression Of ADAM 10 as a Predictor of Lymph Node Metastasis in Oral Squamous Cell Carcinoma-An Immunohistochemical Study.

Lymph node metastasis (LNM) is a well-known prognostic factor in Oral Squamous Cell Carcinoma(OSCC). A biological marker that predicts the Lymph Node Metastasis (LNM) in OSCC cases is the need of the hour. A Disintegrin And Metalloproteinases (ADAMs), a family of proteins that exhibit a metalloproteinase domain play a pivotal role in the pathogenesis of tumor growth and metastasis. This study aims to evaluate whether ADAM 10 can be used as a predictor of lymph node metastasis in OSCC using immunohistochemistry. A total of 90 samples that were categorized into 3 groups were included in the present study. Group I consisted of 30 samples of the normal oral mucosa, and Group II consisted of 30 samples of OSCC without lymph node metastasis. Group III consisted of 30 samples of OSCC with lymph node metastasis. Esophageal Squamous Cell Carcinoma was used as external positive control. Immunohistochemical expression of ADAM10 in their corresponding stained sections was assessed and staining intensity was calculated. ADAM10 immunoreactivity was considered positive when located in cytoplasm or membrane or both. This method is similar to that used by Bamane et al. for OSCC cases. The mean value of the Staining Index score "AxB" was highest in Group III (7.90), followed by Group II (3.13) and least in Group I (0.27). These values were statistically significant. Considering the findings of a higher percentage of ADAM10 positive cells, higher staining intensity, and higher staining index, the overexpression of ADAM10 can be used as an independent marker for OSCC patients to predict the lymph node metastasis.

The effect of endostatin on angiogenesis and osteogenesis of steroid-induced osteonecrosis of the femoral head in a rabbit model.

This study aimed to investigate whether endostatin, a crucial anti-angiogenic factor, plays a negative role in angiogenesis and osteogenesis and aggravates the progression of osteonecrosis of the femoral head induced by steroid use in a rabbit model. 66 New Zealand white rabbits were randomly divided into four groups: glucocorticoid model (GC) group (GC group, n = 18), glucocorticoid model and endostatin group (GC;ES group, n = 18), ES group (ES group, n = 18), and blank control group (CON group, n = 12). In the GC group, 10 μg/ kg lipopolysaccharide (LPS) was intravenously injected into the ear margin, and 24h after LPS injection, 20 mg/kg GC methylprednisolone (MPS) was injected into the gluteus muscle three times, each time at an interval of 24h. The animals of the GC;ES group were given as same treatment as the GC group, except for the addition of ES. MPS was not used in the ES group and CON group. ES group was only given ES, while the CON group was only given the same amount of normal saline. All animals successfully established models of femoral head necrosis, and then the difference among the Immunohistochemistry, Quantitative polymerase chain reaction (qPCR) analysis, Enzyme-linked immunosorbent assay, Biomechanical test, etracyclline-calcein double labeling, and Van Gieson staining indices were compared among the four groups. The combination of MPS and LPS was successful in establishing the femoral head necrosis model in New Zealand white rabbits. The incidence of osteonecrosis after MPS and LPS intervention was 70% (7/10), while that plus ES was 100% (10/10). At the same time, after MPS and LPS intervention, while the empty bone lacuna rate of the femoral head was significantly increased, the number of osteo- blasts was decreased. Also, the expressions of CD31 positive cells, Runx2, Osterix, COL1A1, and VEGF mRNA in the femoral head were decreased, and the levels of osteogenesis-related protein b-ALP, OCN, and angiogenic factor VEGF in the femoral head were decreased. The percentage of the trabecular bone area (%Tb.Ar), trabecular thickness (Tb.Th), trabecular number (Tb.N), labeled perimeter percent (%L.Pm), mineral apposition rate (MAR), and bone formation rate (BFR/BS) in the femoral head after MPs and LPS intervention detected by tetracycline calcein double labeling and Van Gieson staining decreased significantly, except trabecular separation (Tb.Sp) increased significantly. The compressive strength (CS), elastic modulus (EM), and strain energy (SE) of the femoral head examed by biomechanical measurement decreased significantly. All the above changes were more obvious after adding ES intervention. ES mRNA in the femoral head was undifferentiated and increased in the GC, ES, and GC;ES group compared with group CON. This study has revealed that ES can inhibit angiogenesis and osteogenesis in the femoral head and aggravate the occurrence and development of femoral head necrosis. Thus, antiangiogenic factors may play an important role in the pathogenesis of ONFH.

Acupuncture for patients with type 2 diabetes mellitus with dry eye: protocol for a systematic review and meta-analysis

IntroductionThe global incidence of patients with type 2 diabetes mellitus (T2DM) with dry eye is increasing annually, which imposes additional healthcare costs and financial burden on families and societies. In...

Comprehensive Profiling of Tubby-Like Proteins in Soybean and Roles of the GmTLP8 Gene in Abiotic Stress Responses.

Tubby-like proteins (TLPs) are transcription factors that are widely present in eukaryotes and generally participate in growth and developmental processes. Using genome databases, a total of 22 putative TLP genes were identified in the soybean genome, and unevenly distributed across 13 chromosomes. Phylogenetic analysis demonstrated that the predicted GmTLP proteins were divided into five groups (I-V). Gene structure, protein motifs, and conserved domains were analyzed to identify differences and common features among the GmTLPs. A three-dimensional protein model was built to show the typical structure of TLPs. Analysis of publicly available gene expression data showed that GmTLP genes were differentially expressed in response to abiotic stresses. Based on those data, GmTLP8 was selected to further explore the role of TLPs in soybean drought and salt stress responses. GmTLP8 overexpressors had improved tolerance to drought and salt stresses, whereas the opposite was true of GmTLP8-RNAi lines. 3,3-diaminobenzidine and nitro blue tetrazolium staining and physiological indexes also showed that overexpression of GmTLP8 enhanced the tolerance of soybean to drought and salt stresses; in addition, downstream stress-responsive genes were upregulated in response to drought and salt stresses. This study provides new insights into the function of GmTLPs in response to abiotic stresses.