The Role of Laundry Detergents in the Elements of Skin Pathogen

Background: The Corona pandemic fuelled up skin pathogen challenges in young and adults, the antimicrobial efficacy of laundry detergents could be considered particularly. However, no available data focusing on the form of laundry detergent, additives and conditions affect the antimicrobial efficacy. This study simulated washing procedures to investigate the antibacterial and antifungal activity of laundry detergents.Methods and Results: Mimic laundry procedures were performed to treat Gram-positive bacteria, Gram-negative bacteria and fungus, colony counting and propidium iodide staining were used to assess the antimicrobial activity. Powder detergent A, NaBO3*4H2O with the tetraacetylethylenediamine, 2Na2co3.3H2O with tetraacetylethylenediamine could achieve a > 5-log10 reduction of three microbial colony generation. Anionic surfactant sodium dodecylbenzene sulphonate (SDBS) group had the strongest fluorescence intensity in three microbial propidium iodide staining.Conclusions: Powder form laundry detergents are superior to liquid form, peroxide-based bleaches and bleach activator in solid form, the solid surfactants with matched PH and alkyl chain length showed a considerable antimicrobial effect.

High expression of MAPK-14 promoting the death of chondrocytes is an important signal of osteoarthritis process

Background Osteoarthritis (OA) is one of the most common degenerative diseases worldwide. Many researchers are studying the pathogenesis of OA, however, it is still unclear. Methods Screening and validation of OA relevant hub genes are an important part of exploring their potential molecular mechanism. Therefore, this study aims to explore and verify the mechanisms of hub genes in the OA by bioinformatics, qPCR, fluorescence and propidium iodide staining. Results Microarray datasets GSE43923, GSE55457 and GSE12021 were collected in the Gene Expression Omnibus (GEO), including 45 samples, which divided into 23 osteoarthritis knee joint samples and 22 samples of normal knee joint. Thereafter, 265 differentiallyexpressedgenes (DEGs) were identified in all, which divided into 199 upregulated genes and 66 downregulated genes. The hub genes MAPK-14, PTPRC, PTPN12 were upregulated, while B9D1 was downregulated. In order to further confirm the expression of screening differential genes in human chondrocytes, the human chondrocytes were extracted from a joint replacement surgery and stained with toluidine blue for identification. Compared with normal chondrocytes, OA chondrocytes had high expression of COL I protein and low expression of COL II protein. The expression levels of MAPK-14, PTPRC and PTPN12 in OA chondrocytes were significantly higher than the expression levels of B9D1 in normal chondrocytes. Moreover, the inflammatory necrosis of OA chondrocytes was increased compared with the normal chondrocytes by propidium iodide staining. Conclusions The high expression of MAPK-14 works as a promoter of chondrocytes death and an important signal of the osteoarthritis process.

Optimization of the method for isolation of epithelial cells from the non-glandular part of the rat stomach for flow cytometry

Traditional methods in cell proliferation studies are based on immunohistochemical detection of proliferating cells in the target tissue. Since they are time consuming, optimization of novel, more efficient methods is important for large scale proliferation studies. In this study, we aimed to optimize the isolation of single epithelial rat forestomach cells for flow cytometry. As a marker of cellular proliferation we used the Ki-67 antibody to detect this nuclear protein expressed in proliferating cells. We also performed immunohistochemical detection of Ki-67 positive cells and propidium iodide staining to validate the results. 3-tert- butyl -4-hydroxyanisole was used as the positive control to ensure cellular proliferation. The results showed that isolation of epithelial cells with collagenase, trypsin and cell strainer ensures great cell viability (>95%) and the purity of the samples. Flow cytometry and immunostaining with the Ki-67 antibody indicated that 3-tert- butyl-4-hydroxyanisole treatment leads to a significant increase in proliferation. A significant positive correlation was observed between the results obtained by immunohistochemistry and flow cytometry, but the flow cytometric data had a smaller measurement error, suggesting the equal sensitivity and greater accuracy of this method. Propidium iodide staining showed that the percentage of cells in the G2+S phase of the cell cycle correlated positively with the percentage of Ki-67 positive cells assessed by flow cytometry, indicating that Ki-67 positive cells reflect an active dividing cell pool. We conclude that the isolation of forestomach epithelial cells described is a simple and reliable method for obtaining viable cells for use in flow cytometry. Compared to immunohistochemistry, flow cytometric detection of the Ki-67 antigen is equally sensitive, but much faster and provides more accurate results.

Microampere electric currents caused bacterial membrane damage and two-way leakage in short time

Physical agents such as low electric voltages and currents have recently gained attention for antimicrobial treatment due to their bactericidal capability. Although microampere electric currents were shown to suppress the growth of bacteria, it remains unclear to what extent the microampere currents damage bacterial membrane. Here, we investigated the membrane damage and two-way leakage caused by microampere electric currents (≤ 100 μA) in a short time (30 min). Based on MitoTracker staining, propidium iodide staining, filtration assays, and quantitative single-molecule localization microscopy, we found that microampere electric currents caused significant membrane damages and allowed two-way leakages of ions, small molecules and proteins. This study paves the way to new development and antibiotic applications of ultra-low electric voltages and currents.Statement of SignificancePrevious studies showed that treating bacteria with milliampere electric currents for 72 hours led to significant damages of the bacterial membrane. However, it remains unclear to what extent membrane damages and two-way (i.e. inward and outward) leakages are caused by lower electric currents in a shorter time. In this work, we set out to answer this question. We carried out several assays on the bacteria treated by microampere electric currents of ≤ 100 μA for 30 min, including MitoTracker staining, propidium iodide staining, filtration assays, and quantitative single-molecule localization microscopy. We found and quantified that the membrane damages were caused by microampere electric currents in half an hour and allowed two-way leakages of ions, small molecules, and proteins.

The Role of Acetate in the Antagonization of Oxalate: A Potential Causative Molecule for Heart Disease and Cancer Death

Previous reports demonstrated the anticancer effects and the protective effects on cardiovascular diseases of vinegar products. The molecular mechanism underlying these phenomena is not well elucidated. It was proposed that carcinogenesis is triggered by the formation of local strong acids such as HCl. Cancer cells may overproduce weak or moderate organic acids such as oxalate to antagonize strong acids, and calcium oxalate may cause organ failure and death. This study aimed at elucidating the underlying mechanism on the antagonism of apoptosis by acetate on oxalate. Quantitation of cell apoptosis of HEK293T cells in the presence of sodium oxalate and compounds with similar structures to oxalate was conducted by using Annexin V-fluorescein isothiocyanate/propidium iodide staining via flow cytometry. The data indicate that acetate could attenuate the proapoptotic functions of oxalate. This study yields insight into the anticancer and antidisease functions of vinegar products and opens up a new path in the use of weak acetic acid in the prevention and treatment of cancer.

Role of C1q in spleen-derived neutrophil infiltration and neuroinflammation after ICH in mice

Background: Neuroinflammation is a major detrimental role of secondary brain injury after spontaneously intracerebral hemorrhage (ICH). Neutrophil infiltration plays a key role in the pathophysiology of ICH, but the coming resource and mechanism is unknown. This study aims to investigate whether spleen-derived neutrophil infiltration accelerated neuroinflammation and the role of C1q classical pathway.Methods: Male C57 mice were subjected to collagenase-induced ICH. If necessary, splenectomy was performed 2 weeks prior to ICH induction or anti-C1q neutralizing antibody (50mg/kg) was injected intravenously into the tail vein 15 minutes prior. Immunohistochemistry, Propidium Iodide staining, western blotting, ELISA and qRT-PCR were used to study the change of molecular proteins, neuronal cells and inflammatory factors. 7.0T animal MRI was used to assess hydrocephalus.Results: At 0h, 6h, 12h, 24h and 48h post ICH induction, we found a significant increasing tendency of microglia activation and neutrophil infiltration around hematoma and that C1q upregulation was correlated with neuronal decrease, which peaked at 24h after ICH. Here, we demonstrated spleen atrophy and upregulation of neutrophil in spleen 24h after ICH. Splenectomy prior to ICH mice resulted in significant decrease of microglia and neutrophil infiltration compared with that in group of sham-splenectomy. Moreover, both anti-C1q antibody and splenectomy significantly attenuated neutrophil infiltration and neuron death, restored synapse VGAT, alleviated hydrocephalus and inflammatory factors, such as IL-1β, TNF-α and IL-6 after ICH.Conclusion: The study demonstrated that spleen is a major source of brain neutrophil infiltration after ICH. C1q-targeted inhibition of classic complement pathway could prevent spleen-derived neutrophil infiltration and attenuate ICH induced neuroinflammation, which provides a novel therapeutic approach for hemorrhagical stroke.

Cytotoxicity of Triterpene Seco-Acids from Betula pubescens Buds

The present study investigated the magnitude and mechanism of the cytotoxic effect on selected cancer cell lines of 3,4-seco-urs-4(23),20(30)-dien-3-oic acid (1), 3,4-seco-olean-4(24)-en-19-oxo-3-oic acid (2), and 3,4-seco-urs-4(23),20(30)-dien-19-ol-3-oic acid (3) isolated from downy birch (Betula pubescens) buds by carbon dioxide supercritical fluid extraction and gradient column chromatography. Cell viability in six human cancer lines exposed to these compounds was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was quantified by annexin V/propidium iodide staining of gastric cancer AGS and colorectal cancer DLD-1 cells. To evaluate the mechanism of apoptosis, the expression of apoptosis-related proteins was analyzed by Western blot. Compound 1 exhibited non-specific toxicity, while compounds 2 and 3 were specifically toxic to colon and stomach cancer cells. The toxicity of compounds 2 and 3 against these two cell lines was greater than for compound 1. Cleavage of caspase-8, -9, and -3 was found in AGS and DLD-1 cells treated with all three seco-acids, indicating the induction of apoptosis via extrinsic and intrinsic pathways. Therefore, triterpene seco-acids (1–3) decreased cell viability by apoptosis induction. AGS and DLD-1 cells were more susceptible to seco-acids with an oxidized C19 than normal fibroblasts. Hence, it made them a new group of triterpenes with potential anticancer activity.

Molecular hydrogen suppresses free-radical-induced cell death by mitigating fatty acid peroxidation and mitochondrial dysfunction

Molecular hydrogen (H2) was believed to be an inert and nonfunctional molecule in mammalian cells; however, we overturned the concept by reporting the therapeutic effects of H2 against oxidative stress. Subsequently, extensive studies revealed multiple functions of H2 by exhibiting the efficacies of H2 in various animal models and clinical studies. Here, we investigated the effect of H2 on free-radical-induced cytotoxicity using tert-butyl hydroperoxide in a human acute monocytic leukemia cell line, THP-1. Cell membrane permeability was determined using lactate dehydrogenase release assay and Hoechst 33342 and propidium iodide staining. Fatty acid peroxidation and mitochondrial viability were measured using 2 kinds of fluorescent dyes, Liperfluo and C11-BODIPY, and using the alamarBlue assay based on the reduction of resazurin to resorufin by mainly mitochondrial succinate dehydrogenase, respectively. Mitochondrial membrane potential was evaluated using tetramethylrhodamine methyl ester. As a result, H2 protected the cultured cells against the cytotoxic effects induced by tert-butyl hydroperoxide; H2 suppressed cellular fatty acid peroxidation and cell membrane permeability, mitigated the decline in mitochondrial oxidoreductase activity and mitochondrial membrane potential, and protected cells against cell death evaluated using propidium iodide staining. These results suggested that H2 suppresses free-radical-induced cell death through protection against fatty acid peroxidation and mitochondrial dysfunction.

Effect and Mechanism of Survivin on Hypoxia-Induced Multidrug Resistance of Human Laryngeal Carcinoma Cells

This study aimed at clarifying the mechanism and role of survivin in hypoxia-induced multidrug resistance (MDR) of laryngeal carcinoma cells. Human laryngeal cancer cells were incubated under hypoxia or normoxia. The expression of survivin was silenced by performing RNA interference. Additionally, by Western blot and real-time quantitative RT-PCR, survivin expression was detected. The sensitivity of human laryngeal carcinoma cells to multiple drugs was measured by CCK-8 assay. Meanwhile, the apoptosis of cells induced by cisplatin or paclitaxel was assessed by Annexin-V/propidium iodide staining analysis. Under hypoxic conditions, the upregulation of survivin was abolished by RNA interference. Then, CCK-8 analysis demonstrated that the sensitivity to multiple agents of laryngeal carcinoma cells could be increased by inhibiting survivin expression (P<0.05). Moreover, Annexin-V/propidium iodide staining analysis revealed that decreased expression of survivin could evidently increase the apoptosis rate of laryngeal carcinoma cells that were induced by cisplatin or paclitaxel evidently (P<0.05). Our data suggests that hypoxia-elicited survivin may exert a pivotal role in regulating hypoxia-induced MDR of laryngeal cancer cells by preventing the apoptosis of cells induced by chemotherapeutic drug. Thus, blocking survivin expression in human laryngeal carcinoma cells may provide an avenue for gene therapy.