The whitening efficacy of a compound formula examined using an ultraviolet-induced skin melanization model.

BACKGROUND: In Eastern culture, a fair complexion is the standard of beauty, leading to appearance-related distress among women with darker skin or facial pigmentation. Women seek whitening cosmetics to enhance their skin tone or correct their pigmentation, but their safety and effectiveness are paramount factors to consider. In this study, we evaluated the safety and whitening effects of a compound formula denoted as TEST comprising astaxanthin, nicotinamide, arbutin, and tranexamic acid. METHODS: Primary skin irritation and skin-whitening efficacy were examined. Three qualified melanization areas were treated with TEST, 7% ascorbic acid, or a blank. Skin color, the individual type angle (ITA°), and the melanin index (MI) were compared among treatment areas. RESULTS: TEST did not induce a skin response and exhibited a significantly higher ITA° than the blank, while no significant difference was observed with that of 7% ascorbic acid. Furthermore, the MI of TEST was significantly reduced posttreatment. CONCLUSIONS: TEST could be integrated into spot-fading and skin-whitening cosmeceuticals or functional cosmetics.

Promoting public skin health through a national continuing medical education project on cosmetic and dermatologic sciences: a 15-year experience.

Background: The developments in cosmetic sciences and technologies have generated a gap between the cosmetics and their users. Users including regular customers, clinicians, industry personnel, researchers, testing agencies, beauty salon workers, and mass media hardly possess the ability to distinguish truth from falsehood. The gap remained as one major reason for inappropriate cosmetics usage, insufficient efficacy, and even cosmetics adverse reactions (CARs). Methods: Aiming at enhancing the relevant practitioners' cosmetic and dermatologic sciences, we launched a cosmetic and dermatologic sciences continuing medical education (CME) since 2008. The objective of the current study was to evaluate the effectiveness of the CME. We summarized and analyzed the project for the last 15 years. Meanwhile, an online survey consisted of three parts was performed to evaluate the CME and to collect the trainees' comments. Results: A total of 3,923 trainees have participated in the CME project from 2008 to 2022. The trainees included clinicians, industry staffs, biomedical researchers, third-party cosmetics testing staffs, beauty salon staffs, students, and media staffs. The trainees had theory courses on cosmetic and dermatologic sciences, cosmetics DIY practice & video watching, and an optional guided tour during the 4.5-day CME. Eight hundred and twenty-three trainees and 586 control subjects responded to the online survey. The comprehensive test in the second part of the survey demonstrated that compared with the control group, the CME project significantly enhanced the trainees' perception and knowledge regarding the cosmetics formula sciences, basic dermatologic sciences, cosmetics usage, noninvasive measurements, new advances, CARs, and laws (p = 0.000). Trainees of all occupations ranked "basic dermatologic sciences and skin diseases" as the most significant sections. Trainees of all occupations believed the CME has contributed most in "understand the function & efficacy of cosmetics." We noticed the occupational variances. Over 97% of trainees were willing to recommend the CME to the others. Conclusion: The CME project significantly enhanced the trainees' cosmetic and dermatologic sciences, which bridged the gap between cosmetics and public skin health. This multidisciplinary CME also contributed to establishing an interdisciplinary interaction and cooperation platform for the multiple occupations involved in the public skin health maintenance and promotion.

Resveratrol Enhances Temozolomide Efficacy in Glioblastoma Cells through Downregulated MGMT and Negative Regulators-Related STAT3 Inactivation.

Chemoresistance blunts the efficacy of temozolomide (TMZ) in the treatment of glioblastoma (GBM). Elevated levels of O6-methylguanine-DNA methyltransferase (MGMT) and activation of signal transducer and of transcription 3 (STAT3) have been reported to correlate with GBM resistance to alkylator chemotherapy. Resveratrol (Res) inhibits tumor growth and improves drug chemosensitivity by targeting STAT3 signaling. Whether the combined therapy of TMZ and Res could enhance chemosensitivity against GBM cells and the underlying molecular mechanism remains to be determined. In this study, Res was found to effectively improve chemosensitivities of different GBM cells to TMZ, which was evaluated by CCK-8, flow cytometry, and cell migration assay. The combined use of Res and TMZ downregulated STAT3 activity and STAT3-regulated gene products, thus inhibited cell proliferation and migration, as well as induced apoptosis, accompanied by increased levels of its negative regulators: PIAS3, SHP1, SHP2, and SOCS3. More importantly, a combination therapy of Res and TMZ reversed TMZ resistance of LN428 cells, which could be related to decreased MGMT and STAT3 levels. Furthermore, the JAK2-specific inhibitor AG490 was used to demonstrate that a reduced MGMT level was mediated by STAT3 inactivation. Taken together, Res inhibited STAT3 signaling through modulation of PIAS3, SHP1, SHP2, and SOCS3, thereby attenuating tumor growth and increasing sensitivity to TMZ. Therefore, Res is an ideal candidate to be used in TMZ combined chemotherapy for GBM.

Comparative Pharmacokinetics of Scoparone and its Metabolite Scopoletin in Normal and ANIT-induced Intrahepatic Cholestatic Rats.

BACKGROUND: Scoparone, the principal natural active ingredient of Artemisia capillaries (Yin Chen), can effectively treat cholestatic diseases, but the pharmacokinetic properties of scoparone are rarely studied in intrahepatic cholestatic rats. OBJECTIVE: A sensitive and rapid LC-MS/MS method was established to detect scoparone and its metabolite of scopoletin in rat plasma and then compare their plasma pharmacokinetic differences between the normal and ANITinduced cholestasis rats. METHODS: Positive ionization was used to separate scoparone and scopoletin using acetonitrile and 0.1 % formic acid water as the mobile phase on a Hypersil ODS-BP column. RESULTS: The calibration curves presented good linearity (R=0.9983 and 0.9989) in the concentration range of 10- 10000 ng/mL and 0.5-500 ng/mL for scoparone and scopoletin, respectively. The precision of ≤ 9.4% and the accuracy ranged from -6.4% to 6.8% were recorded over three validation runs, and the recovery was higher than 83.9%. Under different storage conditions, scoparone and scopoletin were stable. Therefore, we studied the pharmacokinetic properties of scoparone and scopoletin in rats after a single oral administration with the above method. According to the results, the pharmacokinetic parameters of AUC, t1/2, and Cmax values of scoparone in the ANIT group were increased by 106%, 75%, and 44%, respectively, while these values of scopoletin were increased by 142%, 62%, and 65%. CONCLUSION: The findings indicated that the pharmacokinetic properties of scoparone and scopoletin were significantly different between the normal and ANIT-induced cholestasis rats, which suggested that the clinical application dosage of scoparone should be adjusted according to the liver function of patients.

Treatment of Stretch Marks Using a New Formulation Combining Nanofractional Radiofrequency Plus Magnetic Nanofractional Radiofrequency.

INTRODUCTION: Stretch marks are common atrophic dermal scars with significant physical and psychological effects. Therefore, there is a need for effective cosmetics and procedures for stretch mark treatment. This study aimed to evaluate the efficacy and safety of a novel treatment for stretch marks that is made up of topical formulations containing beta-glucan combined with nanofractional radiofrequency. METHODS: This randomized, blinded control trial enrolled 64 Chinese women aged 20-45 years at > 6 months after delivery with obvious white or silver abdominal stretch marks. Participants were randomly allocated to group A (blank group), group B (topical product group), group C (product combined with nanofractional radiofrequency), and group D (vehicle combined with nanofractional radiofrequency). The stretch mark width, skin elasticity, skin color, skin thickness, and collagen density were noninvasively measured. Two trained assessors evaluated the severity, color, outline, and relaxation of the striae. RESULTS: Group C showed the best treatment efficacy, with no adverse effects observed during the study period. CONCLUSION: Our findings indicate that stretch mark treatment using topical formulations containing beta-glucan, combined with nanofractional radiofrequency plus magnetic nanofractional radiofrequency, is tolerable and effective. TRIAL REGISTRATION: ChiCTR2200056725.

Clinical study of a spray containing birch juice for repairing sensitive skin.

Sensitive skin is described as an unpleasant sensory response to a stimulus that should not cause a sensation. Sensitive skin affects an increasing proportion of the population. Sixty-seven participants who tested positive to lactic acid sting test were recruited and randomized into two groups to observe the clinical efficacy and safety of a new birch juice spray for repairing sensitive skin. One group used test spray A, while the other group used spray B as a control. Both groups were sprayed six times daily for 28 days. Noninvasive testing instruments were used to measure stratum corneum hydration, sebum content, transepidermal water loss rates, skin blood perfusion and current perception threshold before and after using spray. Facial images were captured by VISIA-CR, and the image analysis program (Image-Pro Plus) was used to analyze these to obtain the redness value of the facial skin. Moreover, lactic acid sting test scores and participants' self-assessments were also performed at baseline, week 2 and week 4. Both sprays A and B significantly decreased the lactic acid sting test score, transepidermal water loss rates, skin blood perfusion, and redness, while increasing the stratum corneum hydration. Compared to spray B, spray A increased sensory nerve thresholds at 5 Hz and decreased the transepidermal water loss rates, skin blood perfusion, and lactic acid sting test score. Sprays containing birch juice improved cutaneous biophysical properties in participants with sensitive skin.

Jade material in vitro and in vivo: A study on the anti-inflammatory and repair efficacy of jade material on the skin.

OBJECTIVE: The biological safety of natural jade materials and assembled jade-activated materials on cells and their anti-inflammatory and damage repair functions, as well as the repair function on sensitive skin, were studied utilizing in vitro cell biology and in vivo. METHODS: Human skin fibroblasts were used as model cells to conduct cytotoxicity experiments in vitro, and the effects on the expression of inflammatory factors and growth factor-related genes in fibroblasts were explored. The gene expression values of inflammatory factors IL-1, IL-6, TNF-α and cytokines epidermal growth factors, fibroblast growth factors and COL1A1 in fibroblasts were measured by polymerase chain reaction test. Thirty women with sensitive skin were selected to apply a mask containing jade extract three times a week. After two weeks, non-invasive measures related to skin sensitivity were tested. RESULTS: We confirmed the presence of anti-inflammatory effects in both jade materials, with the effects of the assembled activated jade material being superior to that of the natural jade material. Jade extracts significantly increased the gene expressions of EGF, FGF and COL1A1 in HDF. The results of the in vivo study showed that the mask containing jade extract could significantly increase the skin hydration and decrease the rate of transepidermal water loss and skin lactic acid sting test scores after two weeks of use. Subjective evaluations confirmed improvements in skin dryness, smoothness and fineness. No new sensitization occurred in subjects, and the product was non-irritating. No adverse skin reactions were observed during the test. CONCLUSIONS: The jade materials were able to downregulate the expression of inflammatory factor genes, up-regulate the expression of growth factor genes, and improve the anti-inflammation and repair ability of skin. Furthermore, the test results of participants with sensitive skin after using the mask containing jade extract showed that the mask has repairing ability.

OBJECTIF: L'innocuité du jade naturel, et des substances assemblées activées par le jade, sur les cellules, leurs effets anti-inflammatoire et réparateur, ainsi que leur action réparatrice sur les peaux sensibles ont été étudiés au moyen de la biologie cellulaire in vitro et in vivo. MÉTHODES: Des fibroblastes de peau humaine (HDF - Human Dermal Fibrolasts) ont été utilisés comme cellules modèles pour réaliser des tests de cytotoxicité in vitro, et les effets sur l'expression des facteurs inflammatoires et des gènes associés aux facteurs de croissance dans les fibroblastes ont été étudiés. Les valeurs de l'expression génique des facteurs inflammatoires IL-1, IL-6, TNF-α, des facteurs de croissance épidermique des cytokines (EGF - Epidermal Growth Factor), des facteurs de croissance des fibroblastes (FGF - Fibroblast Growth Factor), et du COL1A1 (Gène Collagen, type I, alpha 1) dans les fibroblastes ont été mesurées au moyen d'un test de réaction en chaîne par polymérase. Trente femmes présentant une peau sensible ont été sélectionnées pour appliquer un masque contenant de l'extrait de jade trois fois par semaine. Au bout de deux semaines, des mesures non invasives de la sensibilité de la peau ont été réalisées. RÉSULTATS: Nous confirmons la présence d'effets anti-inflammatoires pour les deux substances, avec de meilleurs résultats pour la substance assemblée activée par le jade comparé à ceux du jade naturel. Les extraits de jade ont significativement augmenté l'expression de l'EGF, du FGF, et du COL1A1 dans les HDF. Les résultats de l'étude in vivo ont révélé que le masque contenant de l'extrait de jade pouvait améliorer significativement l'hydratation de la peau, réduire le pourcentage de perte en eau trans-épidermique et améliorer les résultats du test de piqûre d'acide lactique (LAST - Lactic Acid Sting Test) de la peau après deux semaines d'utilisation. Les évaluations subjectives ont confirmé des améliorations de la sécheresse cutanée, de la douceur et de la finesse du grain de la peau. Aucune nouvelle sensibilisation n'est apparue chez les sujets, et le produit s'est avéré non-irritant. De même, aucune réaction cutanée indésirable n'a été observée pendant le test. CONCLUSIONS: Le jade a été capable de réguler à la baisse l'expression des gènes associés aux facteurs inflammatoires, de réguler à la hausse l'expression des gènes associés aux facteurs de croissance, et d'améliorer les capacités anti-inflammatoire et réparatrice de la peau. De plus, après utilisation du masque contenant de l'extrait de jade, les résultats des tests chez les participantes ayant une peau sensible ont démontré que ce masque avait une capacité réparatrice.

Increased brain uptake of pterostilbene loaded folate modified micellar delivery system.

Effective chemotherapy for clinical treatment of brain diseases is still lacking due to the poor penetration of the blood-brain barrier (BBB). The aim of this study was to construct a folate modified pterostilbene (Pt) loaded polymeric micellar delivery system (F-Pt/M) with mPEG-PCL as carrier material to aim at penetrating the BBB for brain tissue targeting via receptor-mediated endocytosis. In this study, F-Pt/M was prepared using thin-film hydration method and then optimized by response surface methodology (RSM) with the entrapment efficiency (EE), drug loading (DL) and hydrodynamic diameter (HD) as indexes. The average hydrodynamic diameter and zeta potential of optimal F-Pt/M were 133.2 nm and 24.6 mV, respectively. DL (18.3%) and EE (98.6%) made the solubility of Pt in water about 25 times higher than that of crude Pt. Results of DSC evaluation revealed that drugs were successfully encapsulated inside the polymeric micelles. TEM images showed that homogeneous spherical micellar structures with a narrow size distribution were developed. The release result in vitro showed that F-Pt/M presented sustained release behavior compared to control free Pt solution. Compared to non-targeted Pt/M, F-Pt/M had a significantly higher cytotoxicity against FR-overexpressing A172 cells. In vitro cellular uptake tests illustrated that the micellar delivery system could significantly improve the accumulation of drugs in target cells via receptor-mediated endocytosis. BBB penetration value (P) of F-Pt/M was about 4 folds higher than that of free Pt group. In addition, drug targeting index (DTI) was calculated to determine targeting of F-Pt/M to the brain which was found to be 4.89, implying improved brain targeting was achieved. Hence, the developed F-Pt/M exhibited great potential for delivering more drug molecules across the BBB for the treatment of brain diseases.

Effectiveness of a Radiofrequency Device for Rejuvenation of Aged Skin at Home: A Randomized Split-Face Clinical Trial.

INTRODUCTION: Several techniques, including the use of radiofrequency (RF) devices, are currently utilized for the treatment of skin aging. This study aimed to evaluate the anti-aging effects imparted by a home-based RF beauty device and to compare these results with those of a marketed anti-aging cosmetic in vivo. METHODS: Thirty-three women aged 35-60 years were enrolled in this randomized, controlled, split-face trial. This study involved a 12-week trial with five repeated measurements (at baseline, 2, 4, 8 and 12 weeks). One side of the face was randomly selected to be part of the experimental group and treated with the RF beauty device, while the other side was considered as control and was treated with an anti-aging cosmetic. Treatment safety was evaluated. Skin wrinkles, hydration, radiance, elasticity, color and thickness were evaluated using noninvasive equipment. RESULTS: Thirty-two participants completed the study; one withdrew for personal reasons. Compared with the anti-aging cosmetic-treated facial side, the experimental side showed statistically significant improvements in wrinkles, skin radiance, color and thickness (p < 0.05). CONCLUSIONS: The home-based RF beauty device was safe and effective for rejuvenation. The device was more effective than the commercially available anti-aging cosmetics.

Enhancement of gemcitabine efficacy by K73-03 via epigenetically regulation of miR-421/SPINK1 in gemcitabine resistant pancreatic cancer cells.

BACKGROUND: Gemcitabine (GCB) is a first-line chemotherapeutic drug for pancreatic cancer (PCa). However, the resistance begins developing within weeks of chemotherapy. SPINK1 overexpression enhances resistance to chemotherapy. In a recent study, our laboratory established that the oleanolic acid (OA) derivative, K73-03, had a strong inhibitory effect on a SPINK1 overexpressed PCa cells. PURPOSE: In our current study, we studied the enhancement of GCB inhibitory effect by K73-03, a new novel OA derivative, alone or in combination with GCB on the GCB-resistant PCa cells by mitochondrial damage through regulation of the miR-421/SPINK1. METHODS: We detected the binding between miR-421 and SPINK1-3'-UTR in GCB-resistant PCa cells using Luciferase reporter assays. Cells viability, apoptosis, migration, and mitochondrial damage were investigated. RESULTS: The results demonstrated that the combination of K73-03 and GCB suppressed the growth of AsPC-1 and MIA PaCa-2 cells synergistically, with or without GCB resistance. Mechanistic findings showed that a combination of K73-03 and GCB silences SPINK1 epigenetically by miR-421 up-regulating, which leads to mitochondrial damage and inducing apoptosis in GCB-resistant PCa cells. CONCLUSION: We found an interesting finding that the 73-03 in combination with GCB can improve GCB efficacy and decrease PCa resistance, which induced apoptosis and mitochondrial damage through epigenetic inhibition of SPINK1 transcription by miR-421 up-regulation. This was the first study that used OA derivatives on GCB-resistant PCa cells, so this combined strategy warrants further investigation.

Triptolidenol, isolated from Tripterygium wilfordii, disrupted NF-κB/COX-2 pathway by targeting ATP-binding sites of IKKß in clear cell renal cell carcinoma.

Triptolidenol (TPD) is an epoxy diterpene lactone from Tripterygium wilfordii, which has been used for chronic nephritis in China，and possessed various pharmacological properties, such as anti-inflammatory and anti-cancer activities. However, the precise molecular antitumor mechanism of TPD remains to be elucidated. In this study, we investigated the effects of TPD on human clear cell renal cell carcinoma (ccRCC) and investigated its precise anti-tumor mechanisms. It was showed that TPD significantly suppressed ccRCC cell proliferation, cell migration, and induced cell cycle arrest at S phase. Furthermore, TPD also induced apoptosis by activating the cytochrome c (cyt c)/caspase cascade signaling pathway. Moreover, using confocal immunofluorescence, a dual-luciferase reporter assay and molecular docking study, the results showed that TPD obviously reduced the expression of COX-2 by inhibiting the kinase activity of IKKß via targeting its ATP-binding domain, and then attenuating the transactivation of NF-κB. Collectively, our study demonstrated that TPD suppressed renal cell carcinoma growth through disrupting NF-κB/COX-2 pathway by targeting ATP-binding sites of IKKß, and provided pharmacological evidence that TPD exhibits potential use in the treatment of COX-2-mediated diseases such as ccRCC.

Intra-articular delivery of flurbiprofen sustained release thermogel: improved therapeutic outcome of collagenase II-induced rat knee osteoarthritis.

Knee osteoarthritis (OA) is a common degenerative disease. Intra-articular administration of flurbiprofen is frequently employed in clinic to treat OA, while repeated injections are required because of the limited effective duration. To improve therapeutic outcome and prolong the treatment interval, a poly(ε-caprolactone-co-lactide)-b-poly(ethylene glycol)-b-poly(ε-caprolactone-co-lactide) (PCLA-PEG-PCLA) triblock copolymer based flurbiprofen thermosensitive gel for the sustained intra-articular drug delivery was designed in this study. The anti-OA effects of this flurbiprofen thermogel were investigated on collagenase II-induced rat knee OA model by multiple approaches and compared with that of conventional sodium hyaluronate and flurbiprofen injecta. In vitro drug release studies indicated that flurbiprofen was sustained released from the thermosensitive gel for more than three weeks. This sustained drug release system exerted comparable short-term analgesic effects and distinctly improved long-term analgesic efficacy in terms of the increased percentage of the total ipsilateral paw print intensity and the reduced Knee-Bend scores of OA rats. The inflammatory response was attenuated in the samples of flurbiprofen gel treated group by showing decreased IL-1, IL-6, and IL-11 levels in the joint fluid and down-regulated IL-1, IL-6, IL-11, COX-2, TNF-α, and NF-κB/p65 expression in the articular cartilages. The results suggest the suitability of thermosensitive copolymer PCLA-PEG-PCLA for sustained intra-articular effects of flurbiprofen and provide in vivo experimental evidence for potential clinical application of this flurbiprofen delivery system to better management of OA cases.

Pannexin 1 channels in renin-expressing cells influence renin secretion and blood pressure homeostasis.

Kidney function and blood pressure homeostasis are regulated by purinergic signaling mechanisms. These autocrine/paracrine signaling pathways are initiated by the release of cellular ATP, which influences kidney hemodynamics and steady-state renin secretion from juxtaglomerular cells. However, the mechanism responsible for ATP release that supports tonic inputs to juxtaglomerular cells and regulates renin secretion remains unclear. Pannexin 1 (Panx1) channels localize to both afferent arterioles and juxtaglomerular cells and provide a transmembrane conduit for ATP release and ion permeability in the kidney and the vasculature. We hypothesized that Panx1 channels in renin-expressing cells regulate renin secretion in vivo. Using a renin cell-specific Panx1 knockout model, we found that male Panx1 deficient mice exhibiting a heightened activation of the renin-angiotensin-aldosterone system have markedly increased plasma renin and aldosterone concentrations, and elevated mean arterial pressure with altered peripheral hemodynamics. Following ovariectomy, female mice mirrored the male phenotype. Furthermore, constitutive Panx1 channel activity was observed in As4.1 renin-secreting cells, whereby Panx1 knockdown reduced extracellular ATP accumulation, lowered basal intracellular calcium concentrations and recapitulated a hyper-secretory renin phenotype. Moreover, in response to stress stimuli that lower blood pressure, Panx1-deficient mice exhibited aberrant "renin recruitment" as evidenced by reactivation of renin expression in pre-glomerular arteriolar smooth muscle cells. Thus, renin-cell Panx1 channels suppress renin secretion and influence adaptive renin responses when blood pressure homeostasis is threatened.

Short-term skin reactions following use of N95 respirators and medical masks.

BACKGROUND: In the context of the COVID-19 pandemic, cases of adverse skin reactions related to the wearing of masks have been observed. OBJECTIVES: To analyze the short-term effects of N95 respirators and medical masks, respectively, on skin physiological properties and to report adverse skin reactions caused by the protective equipment. METHODS: This study used a randomized crossover design with repeated measurements. Twenty healthy Chinese volunteers were recruited. Skin parameters were measured on areas covered by the respective masks and on uncovered skin 2 and 4 hours after donning, and 0.5 and 1 hour after removing the masks, including skin hydration, transepidermal water loss (TEWL), erythema, pH, and sebum secretion. Adverse reactions were clinically assessed, and perceived discomfort and non-compliance measured. RESULTS: Skin hydration, TEWL, and pH increased significantly with wearing the protective equipment. Erythema values increased from baseline. Sebum secretion increased both on the covered and uncovered skin with equipment-wearing. There was no significant difference in physiological values between the two types of equipment. More adverse reactions were reported following a N95 mask use than the use of a medical mask, with a higher score of discomfort and non-compliance. CONCLUSIONS: This study demonstrates that skin biophysical characters change as a result of wearing a mask or respirator. N95 respirators were associated with more skin reactions than medical masks.

Endothelial Pannexin 1 Channels Control Inflammation by Regulating Intracellular Calcium.

The proinflammatory cytokine IL-1ß is a significant risk factor in cardiovascular disease that can be targeted to reduce major cardiovascular events. IL-1ß expression and release are tightly controlled by changes in intracellular Ca2+ ([Ca2+]i), which has been associated with ATP release and purinergic signaling. Despite this, the mechanisms that regulate these changes have not been identified. The pannexin 1 (Panx1) channels have canonically been implicated in ATP release, especially during inflammation. We examined Panx1 in human umbilical vein endothelial cells following treatment with the proinflammatory cytokine TNF-α. Analysis by whole transcriptome sequencing and immunoblot identified a dramatic increase in Panx1 mRNA and protein expression that is regulated in an NF-κB-dependent manner. Furthermore, genetic inhibition of Panx1 reduced the expression and release of IL-1ß. We initially hypothesized that increased Panx1-mediated ATP release acted in a paracrine fashion to control cytokine expression. However, our data demonstrate that IL-1ß expression was not altered after direct ATP stimulation in human umbilical vein endothelial cells. Because Panx1 forms a large pore channel, we hypothesized it may permit Ca2+ diffusion into the cell to regulate IL-1ß. High-throughput flow cytometric analysis demonstrated that TNF-α treatments lead to elevated [Ca2+]i, corresponding with Panx1 membrane localization. Genetic or pharmacological inhibition of Panx1 reduced TNF-α-associated increases in [Ca2+]i, blocked phosphorylation of the NF-κB-p65 protein, and reduced IL-1ß transcription. Taken together, the data in our study provide the first evidence, to our knowledge, that [Ca2+]i regulation via the Panx1 channel induces a feed-forward effect on NF-κB to regulate IL-1ß synthesis and release in endothelium during inflammation.

JAK3 inhibitors based on thieno[3,2-d]pyrimidine scaffold: design, synthesis and bioactivity evaluation for the treatment of B-cell lymphoma.

JAK3 is predominantly expressed in hematopoietic cells and has been a promising therapeutic target for the treatment of B-cell lymphoma. In this study, a new class of thieno[3,2-d]pyrimidines harboring acrylamide pharmacophore were synthesized as potent covalent JAK3 inhibitors (IC50 < 10 nM). Among them, 9a and 9 g displayed the strongest inhibitory potency against JAK3 kinase activity, with IC50 values of 1.9 nM and 1.8 nM, respectively. Furthermore, compared with the reference agents, Spebrutinib and Ibrutinib, 9a not only demonstrated enhanced antiproliferative activity against B lymphoma cells, but also showed very weak proliferative inhibition against normal peripheral blood mononuclear cells (PBMCs) at a concentration of 20 µM. Analysis of the mechanism revealed that 9a could induce the obvious apoptosis in B lymphoma cells and prevent JAK3-STAT3 cascade as well as BTK pathway. Taken together, 9a may be served as a potential new JAK3 inhibitor for the treatment of B-cell lymphoma.

Novel Pyrimidines as Multitarget Protein Tyrosine Kinase Inhibitors for the Treatment of Idiopathic Pulmonary Fibrosis (IPF).

A new class of pyrimidine derivatives were identified as potent protein tyrosine kinase (PTK) inhibitors for the treatment of idiopathic pulmonary fibrosis (IPF). Most of these small-molecule inhibitors displayed strong enzymatic activity against BTK and JAK3 kinases at concentrations lower than 10 nM. The representative compound N-(3-((5-chloro-2-(4-((1-morpholino)acetylamino)phenylamino)-4-pyrimidinyl)amino)phenyl)acrylamide (6 a) also exhibited high inhibitory potency toward both BTK and JAK kinase families, as well as ErbB4, at a concentration of 10 nM, achieving rates of inhibition higher than 57 %. Additionally, in vivo biological evaluations showed that 6 a can remarkably decrease the severity of IPF disease. All these investigations suggested that the multi-PTK inhibitor 6 a may serve as a promising agent for the treatment of IPF.

Quercetin and Quercitrin Attenuates the Inflammatory Response and Oxidative Stress in LPS-Induced RAW264.7 Cells: In Vitro Assessment and a Theoretical Model.

BACKGROUND: Nowadays, atmospheric pollutants, ultraviolet rays, and other factors cause the imbalance of cell redox, resulting in skin oxidative damage. There is an interaction between inflammatory response and oxidative stress, which often involve networks of reactions and serve to amplify each other. Quercetin and quercitrin, with strong antioxidant and anti-inflammatory properties, were widely applied in cardiovascular disease, osteoporsis, pulmonary disease, etc. However, the regulation mechanism of quercetin and quercitrin on various inflammatory skin diseases is still not clear. PURPOSE: In this study, quercetin and quercitrin were used to investigate whether they had anti-inflammatory and anti-ROS effects. Besides, theoretical calculation method was also adopted to preliminarily explore the mechanism of the anti-inflammatory and antioxidant effects of these two substances. METHODS: CCK-8 assay was employed to investigate the cytotoxicity. The concentration of NO measured by Griess Reaction System. Moreover, the inflammatory factors (TNF-α, IL-1ß, and IL-6) were reduced in LPS-stimulated RAW264.7 cells were tested by ELISA kits. The trend of ROS changes was detected by DCFH-DA method. Finally, the mechanism of the anti-inflammatory and antioxidant effects of these two substances was carried out by DMol3 package in Materials Studio. RESULTS: CCK-8 assay results guided that the safe concentration of quercetin and quercitrin was lower than 15.0 µg/mL and 22.4 µg/mL, respectively. Also, the concentration of NO could significantly be inhibited by quercetin and quercitrin. Besides, the ELISA results showed that TNF-α, IL-1ß, and IL-6 were reduced in LPS-stimulated RAW264.7 cells after interfering with quercetin and quercitrin. The trend of ROS changes was similar to that of inflammatory factors. Finally, the theoretical calculation illustrated that the oxygen atom on B rings may be the main site of electron cloud density changes, which may suggest a possible mechanism for the anti-inflammatory and ROS scavenging effects of quercetin and quercitrin. CONCLUSIONS: This experiment shows that LPS can induce the overactivating of macrophages and the activated macrophages can subsequently induce inflammatory storms and oxidative stress. Both quercetin and quercitrin can inhibit LPS-induced macrophage inflammation and oxidative stress by experiment and theoretical calculations.

7-O-geranylquercetin contributes to reverse P-gp-mediated adriamycin resistance in breast cancer.

AIMS: To investigate the effect of 7-O-geranylquercetin (GQ), a derivative of quercetin (Q), on reversing drug resistance in breast cancer MCF-7/ADR cells and reveal the mechanisms related to P-glycoprotein (P-gp). MAIN METHODS: Cell viability was determined by MTT assay. Accumulation of adriamycin (ADR) in cells was determined by confocal fluorescence microscope and microplate reader while that of rhodamine (Rh) was measured by flow cytometry. Expression levels of P-gp and MDR1 gene in cells were detected by western blot and Real-Time PCR, respectively. Molecular docking of GQ and Q with P-gp was conducted using AutoDock program. Xenograft model was established by inoculating MCF-7/ADR cells in BALB/c-nude mice. Tumor bearing mice were administered with ADR via tail vein injection and/or GQ (Q) by gavage. Expression levels of P-gp in tissues were detected by western blot and immunohistochemistry. KEY FINDINGS: GQ could reverse drug resistance of MCF-7/ADR cells to ADR. GQ inhibited the efflux of ADR by down-regulating the expression of P-gp protein and its encoding gene MDR1 in MCF-7/ADR cells. Molecular modeling showed that GQ matched with P-gp better than Q. GQ enhanced the antitumor effects of ADR and decreased the expression of P-gp in mice and its activities were higher than that of Q. GQ could reverse drug resistance of MCF-7/ADR cells by down-regulating the expression of P-gp in vitro and in vivo. SIGNIFICANCES: The reversal effect of GQ on P-gp-mediated drug resistance indicates its potential as a reversal agent for drug resistance in cancer chemotherapy.

Negative regulators of STAT3 signaling pathway in cancers.

STAT3 is the most ubiquitous member of the STAT family and involved in many biological processes, such as cell proliferation, differentiation, and apoptosis. Mounting evidence has revealed that STAT3 is aberrantly activated in many malignant tumors and plays a critical role in cancer progression. STAT3 is usually regarded as an effective molecular target for cancer treatment, and abolishing the STAT3 activity may diminish tumor growth and metastasis. Recent studies have shown that negative regulators of STAT3 signaling such as PIAS, SOCS, and PTP, can effectively retard tumor progression. However, PIAS, SOCS, and PTP have also been reported to correlate with tumor malignancy, and their biological function in tumorigenesis and antitumor therapy are somewhat controversial. In this review, we summarize actual knowledge on the negative regulators of STAT3 in tumors, and focus on the potential role of PIAS, SOCS, and PTP in cancer treatment. Furthermore, we also outline the STAT3 inhibitors that have entered clinical trials. Targeting STAT3 seems to be a promising strategy in cancer therapy.