Stemness Signature Predicts Outcomes and Identifies Candidates for Targeted Therapy in Diffuse Large B-Cell Lymphoma

Background: Mounting studies have shown that tumor cells have stem cell-like phenotypes called stemness. The stemness on prognosis and the immunological properties of diffuse large B-cell lymphoma (DLBCL) has not been systematically explored. This study aimed to explore the relationship between stemness, prognosis and immune microenvironment and further to identified potential therapeutic target related to stemness. Methods: The gene expression and clinical information of 1398 diffuse large B-cell lymphoma sample were obtained to calculated stemness based on the single-sample gene set enrichment analysis (ssGSEA). Survival analysis was performed for stemness phenotypes and the association of stemness with clinical features was assessed further. Weighted gene co-expression network analysis (WGCNA) to screen the relevant key genes systematically for stemness of DLBCL. Cell cycle assay and apoptosis assay were used to evaluate the role of cell division cycle 7 (CDC7) inhibitors in DLBCL cell lines. Results: Stemness was significantly associated with overall survival (OS) and progress free survival (PFS) of DLBCL patients. Notably, prolonged OS and PFS was observed in low stemness groups compared to high group, and the international prognostic index (IPI) with a high risk had higher stemness. Furthermore, DLBCL stemness were associated with tumor purity, immune score and stroma score. From stemness module, 16 genes in the co-expression were chosen as key genes and further identified CDC7 as candidate target gene for antitumor. CDC7 inhibitors involving in dequalinium chloride which was approved by Food and Drug Administration (FDA) for anti-microbial and simurosertib which in a phase I clinical trial for multiple solid tumors show antitumor in DLBCL cell line of SU-DHL-2 and SU-DHL-10. Conclusion: In a word, the study integrated DLBCL stemness, clinical characteristics and immunophenotyping, providing an idea of targeting for cold tumors of DLBCL, and also verified the efficacy of inhibitors of key gene of stemness CDC7 in DLBCL. It is significant to develop novel therapeutic targets for diffuse large B cell lymphoma. Figure Legends: Figure (A-B) An overview of the association between known clinical and molecular features (IPI, Hams, gender, and stage) and stemness index in DLBCL cohort. Columns represent samples sorted by stemness index from low to high (top row). Rows represent known clinical and molecular. (C-D) Overall survival and progression free survival of stemness index in DLBCL patients of GSE117556 by Kaplan-Meier analysis. (E-F) Overall survival and progression free survival of stemness index in DLBCL patients of GSE31312 by Kaplan-Meier analysis. (G-I) Boxplots of tumor purity, immune score, stromal score in individual samples stratified by stemness index. (J) WGCNA analysis of differentially expressed genes in DLBCL. Different colors correspond to related modules. (K) Correlation coefficient between the modules and clinical traits with stemness. P-values are listed in the modules. (L-N) Scatter plot analysis of different modules in the MEblue, MEbrown, and MEturquoise modules. (O) Signor analysis of 16 key genes in DisNor. (P) K-M curves of the expression of CDC7 for the DLBCL patients. (Q) Boxplots of the expression of CDC7 in individual samples stratified by Hams. (R) Apoptosis ratio of SU-DHL-10 cells were treated with dequalinium chloride and simurosertib for 48 h. Data are mean ± SD of triplicate experiments. For dequalinium chloride and simurosertib compared with control an unpairedt-test was used. (S) Cell cycle of SU-DHL-10 cells were treated with dequalinium chloride and simurosertib for 48 h. Data are mean ± SD of triplicate experiments. For dequalinium chloride and simurosertib compared with control an unpairedt-test was used. (T) Cell viability of SU-DHL-2 and SU-DHL-10 cells treated with dequalinium chloride and simurosertib at the IC50 concentrations. * p<0.05. ** p<0.01, *** p<0.001. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Evaluation of the effectiveness of dequalinium chloride vaginal tablets in aerobic vaginitis: A placebo-controlled study

Background: Aerobic vaginitis is a disturbance of vaginal homeostasis caused by the colonization of enteric bacteria. They are not clinically well-defined and do not have standard treatment regimes. Hence, the present study was conducted to evaluate the effectiveness of dequalinium chloride topical vaginal administration in aerobic vaginitis in a placebo-controlled manner. Materials and Methods: This study was conducted in a placebo-controlled manner to demonstrate the effectiveness of vaginal 10 mg dequalinium chloride. A total of 30 patients with different vaginal infections were included in the study. The patients included in the study were divided between the drug group and the placebo group and were treated with one vaginal tablet daily for 6 days. The total symptom score, which consists of the assessment of discharge, itching, and burning sensation, and the lactobacillus grade evaluated microscopically, was determined. Results: It was found that the effectiveness of dequalinium chloride was very high compared to placebo (92% vs. 0%). The reproduction frequency in the first visit was statistically significantly lower in the drug group compared to the placebo group (P < 0.001). Conclusion: It was found that dequalinium chloride is effective for the treatment of aerobic vaginitis. However, studies containing a larger sample group, including the long-term effects (efficacy and side effects) of the drug, should be conducted to prove our conclusion.

Application Enzymatic Method for Analysis Vaginal Tabletas “Fluomizin”

Introduction: Development of a new enzymatic kinetic-photometric method for analysis of Dequalinium chloride, based on the use of enzymatic hydrolysis of acetylcholine, for determine the amount of Dequalinium chloride in the vaginal tablets “Fluomizin” No 6. Method: The amount of Dequalinium chloride (DCh) was determined by the degree of inhibition of the enzymatic reaction, which was evaluated by the residual unreacted substrate - acetylcholine. Determination of the residual amount of acetylcholine in the reaction mixture was performed by a kinetic-photometric method using an indicator oxidation reaction of p-phenetidine with peracetic acid, which is formed during the auxiliary reaction of perhydrolysis with addition of excess hydrogen peroxide in the reaction mixture over a period of time. Results: Optimal conditions were determined: pH 8.35 was selected, influence of nature of buffer solution, concentration of acetylcholine (0.05 mg / ml), cholinesterase (0.4 mg / ml), hydrogen peroxide (10,0%) and p-phenethedine (1 %), the incubation time (10 min). Conclusions: The procedure for determination of Dequalinium chloride content in the vaginal tablets “Fluomizin” No 6 was developed. The kinetic parameters Km (Michaelis–Menten constant) and Vmax (maximal velocity) were determined.

Clinical case: patient treated for vulvovaginal candidiasis

Case study of acute vulvovaginal candidiasis treated with medication containing dequalinium chloride as an active ingredient has been reported. Patient A. aged 26 complaining of vaginal and perineal burning, itching, as well as of profuse white, whitish vaginal discharge, was under observation. Vulvovaginal candidiasis was diagnosed based on the laboratory tests. After verifying the diagnosis the patient was prescribed a medication containing dequalinium chloride, with beneficial effect. No vaginal dysbiosis was reported after exposure to medication.

Dequalinium Chloride Effectively Disrupts Bacterial Vaginosis (BV) Gardnerella spp. Biofilms

Bacterial vaginosis (BV) is the most frequent vaginal infection worldwide. It is caused by the overgrowth of anaerobic vaginal pathogens such as Gardnerella spp. BV has been associated with the occurrence of dense multispecies biofilms on the vaginal mucosa. Treatment of biofilm-associated infections such as BV is challenging. In this study, we have tested the role of a quaternary ammonium compound, dequalinium chloride (DQC), in the eradication of Gardnerella spp. biofilms. The effects of the test substance on the biomass and the metabolic activity of the biofilm of Gardnerella spp. were assessed in vitro using a microtiter plate assay. In addition, the effect of DQC on the Gardnerella spp. biofilm was further assessed by using scanning electron microscopy and confocal laser scanning microscopy. The results showed that DQC was particularly effective in the destruction of BV-associated Gardnerella spp. biotypes, impacting both their biomass and metabolic activity. In addition, the disruption of biofilm architecture was evident and was probably caused by multiple mechanisms of action. We conclude that DQC is an antibiofilm agent and is able to efficiently destroy Gardnerella spp. BV-associated biofilms. Therefore, it is a valid option for BV therapy and has the potential to prevent BV recurrences.

Dequalinium-derived nanoconstructs: A promising vehicle for mitochondrial targeting

The power house of the cell; mitochondrion, is a vital organelle for drug targeting in the treatment of many diseases owing to its fundamental duties and function related to cell proliferation and death. The mitochondrial membrane comprises bilayer artifact and pose extremely negative potential which create hurdle for therapeutic molecules in reaching mitochondria. To accomplish mitochondrial targeting, the scientific community has explored diverse pharmaceutical formulations like liposomes, polymeric nanoparticles (NPs), and inorganic NPs. However, the game changing technology was modification of these carriers by mitochondriotropic moiety, dequalinium chloride (DQA) or delivering the chemotherapeutics by DQAsomes. The DQA represents a distinctive mitochondriotropic delocalized cation that display their selectivity towards accumulation in mitochondria of carcinoma cell. Attributed to this characteristics, DQAsomes have been formulated using DQA and explored for successful mitochondrial targeting of bioactives. In this review, we have discussed the effectiveness of DQA nanocarriers which efficiently and selectively transmit the cytotoxic drug to the tumor cell. The DQA based nanoformulations have evidently displayed augmented pharmacological and therapeutic outcome than their counterparts both in vitro and in vivo. Thus, DQAsomes symbolizes an ideal carrier with excellent potential as mitochondial targeting agent.

Optimization of methods for correcting disorders of vaginal normocenosis in the primary treatment of patients

Objective. To study the efficacy and feasibility of using dequalinium chloride during the primary treatment of patients of reproductive age with the symptoms of disorders of vaginal normocenosis. Patients and methods. In 60 patients of reproductive age (31.43 ± 6.44 years), an observational study was conducted, in which the initial data characterizing the state of vaginal microbiota were evaluated, and also the dynamics of their change immediately after treatment and after one month were presented. The analysis of risk factors for vaginal normocenosis disorders and dynamics of changes in the clinical picture amid dequalinium chloride therapy (Fluomizin®) was performed. Results. Significant risk factors for disorders of vaginal normocenosis are: early sexual activity (28.3%), the number of sexual partners (4.32 ± 4.11), their frequent change (13.33%), which determine hormonal and immune changes in the mucosa against the background of endocrine pathology (hypothyroidism in 8.33%) and genital endometriosis (in 35.0%). The prescription of Fluomizin® led to the relief of symptoms (itching, burning, discomfort, discharge and hyperemia) after 6 days of its use, which is confirmed by the data of the study (vaginal swabs, Femoflor 16). Conclusion. Dequalinium chloride (Fluomizin®) can be used to correct the disorders of vaginal normocenosis in the initial treatment of patients before obtaining the results of laboratory studies. Key words: reproductive period, vaginal health, dequalinium chloride, treatment quality