(searched for: doi:10.1016/j.jep.2017.09.031)
Journal of Herbal Medicine, Volume 34; https://doi.org/10.1016/j.hermed.2022.100573
Molecules, Volume 26; https://doi.org/10.3390/molecules26113189
Uncaria tomentosa (Willd.) DC is a woody climber species originating from South and Central America that has been used in the therapy of asthma, rheumatism, hypertension, and blood purification. Our previous study showed that U. tomentosa extracts altered human erythrocyte shape, which could be due to incorporation of the compounds contained in extracts into the erythrocyte membrane. The aim of the present study was to determine how the compounds contained in U. tomentosa extracts incorporate into the human erythrocyte membrane. The study has assessed the effect of aqueous and ethanolic extracts from leaves and bark of U. tomentosa on the osmotic resistance of the human erythrocyte, the viscosity of erythrocyte interior, and the fluidity of erythrocyte plasma membrane. Human erythrocytes were incubated with the studied extracts in the concentrations of 100, 250, and 500 µg/mL for 2, 5, and 24 h. All extracts tested caused a decrease in erythrocyte membrane fluidity and increased erythrocyte osmotic sensitivity. The ethanolic extracts from the bark and leaves increased viscosity of the erythrocytes. The largest changes in the studied parameters were observed in the cells incubated with bark ethanolic extract. We consider that the compounds from U. tomentosa extracts mainly build into the outer, hydrophilic monolayer of the erythrocyte membrane, thus protecting the erythrocytes against the adverse effects of oxidative stress.
Published: 29 January 2021
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Phytomedicine, Volume 79; https://doi.org/10.1016/j.phymed.2020.153327
Frontiers in Oncology, Volume 10; https://doi.org/10.3389/fonc.2020.563779
Many similar characteristics in human and dog cancers including, spontaneous development, clinical presentation, tumor heterogeneity, disease progression, and response to standard therapies have promoted the approval of this comparative model as an alternative to mice. Breast cancer represents the second most frequent neoplasm in humans after lung cancer. Triple-negative breast cancers (TNBC) constitute around 15% of all cases of breast cancer and do not express estrogen receptor (ER), progesterone receptor (PR), and do not overexpress human epidermal growth factor receptor 2 (HER2). As a result, they do not benefit from hormonal or trastuzumab-based therapy. Patients with TNBC have worse overall survival than patients with non-TNBC. Lehmann and collaborators described six different molecular subtypes of TNBC which further demonstrated its transcriptional heterogeneity. This six TNBC subtype classification has therapeutic implications. Breast cancer is the second most frequent neoplasm in sexually intact female dogs after skin cancer. Canine mammary tumors are a naturally occurring heterogeneous group of cancers that have several features in common with human breast cancer (HBC). These similarities include etiology, signaling pathway activation, and histological classification. Molecularly CMTs are more like TNBCs, and therefore dogs are powerful spontaneous models of cancer to test new therapeutic approaches, particularly for human TNBCs. More malignant tumors of the breast are more often ER and PR negative in both humans and dogs. Promising breast cancer biomarkers in both humans and canines are cancer-associated stroma (CAS), circulating tumor cells and tumor DNA (ctDNA), exosomes and miRNAs, and metabolites.
Fitoterapia, Volume 147; https://doi.org/10.1016/j.fitote.2020.104772
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Journal of Ethnopharmacology, Volume 267; https://doi.org/10.1016/j.jep.2020.113494
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Natural Product Research pp 1-6; https://doi.org/10.1080/14786419.2020.1795856
One novel pentacyclic triterpene, 24-dimethoxymethyl-3β,6β,19α- trihydroxy −12-en-28-oic acid (1), along with six known compounds 2–7, were isolated from the canes of Uncaria sessilifructus Roxb. Their structures were determined according to spectroscopic and spectrometric analysis. The anti-inflammatory activities of the isolated compounds (1–7) were scanned against NO production in LPS-activated RAW 264.7 macrophages by MTS assay, however no activities were observed.
Oncology Letters, Volume 20, pp 1111-1118; https://doi.org/10.3892/ol.2020.11666
Although ring finger protein 2 (RNF2) serves an important role in the occurrence, development and regulation of various types of cancer, RNF2 expression in skin squamous cell carcinoma (SCC) remains unknown. The aim of the present study was to investigate the role of RNF2 expression in SCC and adjacent tissues from patients. The protein and gene expression levels of RNF2 in SCC and adjacent tissues were detected by immunohistochemistry (IHC), western blot analysis and semi‑quantitative reverse transcription (RT) PCR. Single factor analysis was used to study the association between RNF2 expression level and the clinicopathological characteristics of patients with SCC. Multifactor Cox survival analysis was used to examine the association between RNF2 expression and the overall survival rate of postoperative patients with SCC. The results from IHC staining demonstrated that the positive expression rate of RNF2 was 84.68% (210/248) and 56.05% (139/248) in SCC and in adjacent tissues, respectively. Furthermore, results from western blot analysis demonstrated that RNF2 protein expression in SCC tissues was significantly higher compared with that in the adjacent tissues (P Introduction Skin squamous cell carcinoma (SCC) is one of the commonest non-melanoma skin cancers worldwide (1,2). SCC is a type of malignant tumor originating from epidermis or adnexal keratinocytes, which often manifests in light exposed areas, such as the scalp, the face and the dorsum of the hand. In Europe and the United States, the incidence rate of SCC ranks second and increases yearly, preceding basal cell carcinoma (3). In China, the incidence rate of SCC is the highest among non-melanoma skin tumors (29.4%) and is slightly higher than that of basal cell carcinoma (28%) (4). At present, the incidence rate of SCC increases annually by 2.6% with the increased population age in China, and 10–15% patients develop metastasis (5,6). Once metastasis occurs or when the disease develops rapidly, the follow-up treatment becomes less efficient and the prognosis of patients is poor. At present, surgical resection remains the best clinical treatment for SCC, followed by radiotherapy and chemotherapy (7). Skin is constantly exposed to chemical stress and ultraviolet radiation, which lead to the overproduction of reactive oxygen species (ROS) (8). Increased oxidative stress can deplete and destroy the skin's non-enzymatic and enzymatic antioxidant defense systems, leading to increased oxidative stress and photosensitivity (9). Increased oxidative stress induces protein oxidation and lipid peroxidation, leading to numerous physiological dysfunctions, including tumor formation and aging (10). Ring finger protein 2 (RNF2), a member of the polycomb genes family, is an ubiquitinated ligase with finger ring structure (11,12). As one of the core members of the polycomb family, RNF2 gene is highly expressed in a variety of human tumors (13–15), and its expression is associated with the degree of malignancy and the prognosis of patients. In addition, RNF2 expression was reported to promote tumor growth and metastasis (16). Bosch et al (13) demonstrated that RNF2 expression in breast cancer tissues is significantly increased compared with normal breast tissues and promotes breast cancer cell invasive ability. Chen et al (14) reported that RNF2 knockdown inhibits the proliferation and invasive ability of pancreatic cancer cells. Li et al (15) demonstrated that RNF10 may be the central target for the regulation of diabetic vascular remodeling due to its anti-hyperproliferative, pro-apoptotic anti-inflammatory activities. However, RNF2 expression in SCC remains unknown. Investigating the association between RNF2 expression and the occurrence and development of SCC may therefore provide novel insights for diagnosis, treatment option and prevention of SCC. The present study aimed to evaluate the role of RNF2 expression in SCC by using immunohistochemistry (IHC), western blot analysis and semi-quantitative reverse transcription (RT) PCR, in order to determine a potential prognostic and diagnostic biomarker for patients with SCC. Materials and methods Patients and controls The present study retrospectively analyzed a cross-section of 248 eligible patients with SCC from a total of 320 patients from the First People's Hospital of Yancheng City, The Fifth People's Hospital of Wuxi, Linyi Traditional Hospital and the First Affiliated Hospital of Zhejiang Chinese Medical University between January 2005 and January 2019. The flow chart for patients screening is presented in Fig. 1. All patients underwent surgical resection. The inclusion criteria were as follows: i) Clinicopathological diagnosis of SCC; ii) no radiotherapy or chemotherapy was administered prior to surgery; iii) clinicopathological and follow-up data were complete; and iv) the location was exposed to at least one of the following: Head, face and neck. The exclusion criteria were as follows: i) Exposure to arsenic; ii) prior-use of immunosuppressive agents; iii) non-fatal or unexplained deaths; iv) incomplete clinicopathological and follow-up data; and v) patients lost to follow up. Figure 1. Screening flow chart for patients with SCC patients. SCC, skin squamous cell carcinoma. The 248 patients with SCC included 140 men and 108 women (age range, 25–86 years; mean age, 57 years). The tumor locations were as follows: 119 samples from the head, 77 samples from the face and 52 cases from the neck. Furthermore, the tumor size was 5 cm for 144 cases. A total of 190 cases exhibited poor differentiation and 58 cases showed a high-to-moderate differentiation. According to Broders' pathological grading criteria for SCC (17), 68 cases were grades I and II and 180 cases were grades III and IV. Adjacent tissue specimens were collected by surgical resection from 248 individuals to serve as a control group (age range, 27–81 years; mean age, 55 years). The follow-up results from the 248 patients enrolled in the present study were obtained via medical records (laboratory data, imaging data and clinical characteristics of the patient during hospitalization) and telephone interviews (follow-up data after discharge: Including survival time, treatment and laboratory data.) Postoperative follow-up was performed every three months during the first year, every four months during the second year, every six months during the third year and every eight months until patient succumbed to the disease. All participants provided oral informed consent. The present study was approved by the Ethics Committee of the First People's Hospital of Yancheng City (approval no. 2017044) and of the Fifth People's Hospital of Wuxi [approval no. HMU (Ethics) 2017-k-133]. IHC IHC was used to detect RNF2 distribution by using the Envision and DAB chromogenic reagent kit (cat. no. MAX-002 MAX007TM). Sections were fixed with 4% paraformaldehyde for 24 h at room temperature. The dehydration was then carried out under a gradually increasing ethanol gradient at room temperature. Tissues were immersed in 70% ethanol, 80% ethanol, 95% ethanol I, 95% ethanol II, anhydrous ethanol I, anhydrous ethanol II and anhydrous ethanol III for 1 h each. Xylene was subsequently added to make the tissues transparent, then immersed in paraffin and finally embedded into wax blocks. Sections were embedded in paraffin and cut into 5-µm-thick sections. Subsequently, sections were dewaxed with xylene I, xylene II and xylene III for 10 min at room temperature, respectively, and then immersed in anhydrous ethanol I, anhydrous ethanol II respectively for 2 min at room temperature, and rinsed with distilled water three times (30 sec/wash). Antigen repair was performed with 0.01 citrate buffer, pH 6.0 at 90°C for 1 h, and incubated with 0.3% hydrogen peroxide (Sigma-Aldrich; Merck KGaA) at room temperature for 30 min to inhibit peroxidase activity. Sections were incubated with the primary rabbit anti-human RNF2 monoclonal antibody (1:100; cat. no. EPR12245; Upstate, Biotechnology, Inc.) overnight at 4°C, washed 3 times with PBS solution (PH 7.4, 1:100v/v; 3 min/wash) and then incubated with polymer-HRP secondary antibody (1:200; cat. no. KXX0022; Dako; Agilent Technologies, Inc.) for 20 min at room temperature. The sections were washed 3 times with PBS solution (3 min/wash) and subsequently incubated with DAB solution (OriGene Technologies, Inc.) at room temperature for 5 min. Tissues were subsequently stained with Improved Harris Hematoxylin Dye Solution (2 g hematoxylin, 5 g aluminum sulfate, 0.2 g potassium iodate, 250 ml 95% ethanol, 750 ml distilled water, 50.0 ml glycerol and 0.3 g citric acid; BASO Biotechnology Co., Ltd.; http://www.baso.com.cn) for 20 sec at room temperature. The positive staining was observed by using a light microscope (magnification, ×400) and the proportion of positive area was calculated. IHC staining scores were determined by two blinded independent pathologists. The positively stained cells in SCC tissues and adjacent tissues were observed in 10 randomly selected high-power fields of view and 100 tumor cells were counted. RNF2 was located in the nucleus under light microscopy. The staining score was determined as follows: Negative (−), no brown-yellow positive staining in tumors or glandular epithelial cells; weakly positive (+), 50% positive cells. For statistical analysis, negative (−) or weakly positive (+) were defined as low expression, whereas positive (++) or strong positive (+++) were defined as high expression. Detection of RNF2 expression by western blot analysis Tissue samples were homogenized, lysed with tissue lysate RIPA lysis buffer (Beijing Solarbio Science & Technology Co., Ltd.), centrifuged at 12,000 × g for 5 min at 4°C, and the supernatan t containing the total proteins was collected. Protein concentration was determined using the BCA kit (Pierce; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. A total of 20 µg protein/lane was separated via SDS-PAGE on a 10% gel and electrotransferred onto polyvinylidene difluoride membranes for 1–2 h. Membranes were blocked with 5% skimmed milk for 1 h at room temperature, and incubated with the following antibodies: Rabbit RNF2 (1:2,000; cat. no. ab101273; Abcam) and rabbit β-actin (1:10,000; cat. no. BS1002; Bioworld Technology, Inc.) overnight at 4°C, followed by incubation with anti-rabbit IgG secondary antibody (1:2,000; cat. no. ab205718; Abcam) at room temperature for 1–2 h. The membranes were analyzed using the Dolphin-Doc Plus gel imaging system (25×25 cm; Wealtec Corp.). Detection of RNF2 mRNA expression levels by RT-CR Total RNA was isolated from tissues using TRIzol® and quantified by Nanodrop spectrophotometer. Total RNA (10 µg/sample) was isolated and used to generate complementary DNA. cDNA was amplified by semi-quantitative PCR and normalized to the internal reference gene β-actin. The sequences of primers used were as follows: RNF2, forward 5′-AGCACAATAATCAGCAAGCACTC-3′, reverse 5′-GCTCCACTACCATTTTCAATCTG-3′; and β-actin, forward 5′-TGGCATCCACGAAACTACC3-3′ and reverse 5′-GTGTTGGCGTACAGGTCTT-3′. The thermocycling conditions were as follows: Pre-denaturation at 95°C for 30 sec, 95°C for 5 sec and 60°C for 30 sec, 40 cycles, and final extension at 60°C for 30 sec. Amplification of RNF2 by PCR was examined by 1.0% agarose gel electrophoresis using a Quantity-One electrophoresis apparatus. The absorbance (a) value of the belt and the reference were read, and the results were expressed by the ratio (sample value/reference value). If the ratio of the SCC value was greater than the reference value, it was positively expressed. Otherwise, it was negatively expressed. Statistical analysis SPSS 13.0 (SPSS, Inc.) software was used for statistical analysis. The χ2 test was used to compare the association between the expression status of RNF2 mRNA and protein levels and the clinicopathological characteristics of patients in adjacent and cancer tissues. Kaplan-Meier survival analysis was used to determine the disease-specific and disease-free survival rates. The log-rank test was used to analyze the difference in survival curves. Multivariable regression analysis was performed to determine the prognostic factors by using the Cox proportional-hazards model. P<0.05 was considered to indicate a statistically significant difference. Results RNF2 protein expression in SCC and adjacent tissues IHC was used to detect the expression of RNF2 in SCC tissues and adjacent tissues. The results demonstrated that RNF2 was mainly expressed in the nucleus of SCC tissue cells. A small amount of RNF2 was found in the cytoplasm (Fig. 2A-D). Furthermore, RNF2 expression in SCC tissues was significantly higher compared with adjacent tissues (P<0.05; Fig. 2). The positive expression rate of RNF2 protein was 84.68% (210/248) and 56.05% (139/248) in SCC tissues and in adjacent tissues, respectively. The results from western blot analysis also revealed that RNF2 expression in SCC tissues was significantly higher compared with that in the adjacent tissues (P<0.05; Fig. 3A-B).
Natural Product Research, Volume 35, pp 4550-4559; https://doi.org/10.1080/14786419.2019.1698572
Lepidium meyenii (Maca) is an edible root plant that grows in the Andean region of Peru. For centuries, the plant has been used as a dietary supplement for its nutritional and therapeutic properties. Maca are rich in high value nutritional elements and secondary metabolites (macaridine, macamides and glucosinolates) with high biological activity. Several studies demonstrated various biological effects of Maca mainly in the field of fertility. The aim of this review is to summarize the state of knowledge on the properties of Maca on male reproduction. Literature data was performed in PubMed with researches published from 2000 to 2019. The research showed results related to the effects of Maca on the quality and quantity of the semen, sexual behaviour and disorders of the male genital tract. Despite the numerous studies carried out on different animal species, further research is needed to clarify the mechanisms of action of Maca. Graphical Abstract
Pharmaceuticals, Volume 12; https://doi.org/10.3390/ph12020072
Up until the first half of the 20th century, silver found significant employment in medical applications, particularly in the healing of open wounds, thanks to its antibacterial and antifungal properties. Wound repair is a complex and dynamic biological process regulated by several pathways that cooperate to restore tissue integrity and homeostasis. To facilitate healing, injuries need to be promptly treated. Recently, the interest in alternatives to antibiotics has been raised given the widespread phenomenon of antibiotic resistance. Among these alternatives, the use of silver appears to be a valid option, so a resurgence in its use has been recently observed. In particular, in contrast to ionic silver, colloidal silver, a suspension of metallic silver particles, shows antibacterial activity displaying less or no toxicity. However, the human health risks associated with exposure to silver nanoparticles (NP) appear to be conflicted, and some studies have suggested that it could be toxic in different cellular contexts. These potentially harmful effects of silver NP depend on various parameters including NP size, which commonly range from 1 to 100 nm. In this study, we analyzed the effect of a colloidal silver preparation composed of very small and homogeneous nanoparticles of 0.62 nm size, smaller than those previously tested. We found no adverse effect on the cell proliferation of HaCaT cells, even at high NP concentration. Time-lapse microscopy and indirect immunofluorescence experiments demonstrated that this preparation of colloidal silver strongly increased cell migration, re-modeled the cytoskeleton, and caused recruitment of E-cadherin at cell-cell junctions of human cultured keratinocytes.
Genes, Volume 9; https://doi.org/10.3390/genes9100513
The prototype cold-shock Y-box binding protein 1 (YB-1) is a multifunctional protein that regulates a variety of fundamental biological processes including cell proliferation and migration, DNA damage, matrix protein synthesis and chemotaxis. The plethora of functions assigned to YB-1 is strictly dependent on its subcellular localization. In resting cells, YB-1 localizes to cytoplasm where it is a component of messenger ribonucleoprotein particles. Under stress conditions, YB-1 contributes to the formation of stress granules (SGs), cytoplasmic foci where untranslated messenger RNAs (mRNAs) are sorted or processed for reinitiation, degradation, or packaging into ribonucleoprotein particles (mRNPs). Following DNA damage, YB-1 translocates to the nucleus and participates in DNA repair thereby enhancing cell survival. Recent data show that YB-1 can also be secreted and YB-1-derived polypeptides are found in plasma of patients with sepsis and malignancies. Here we show that in response to oxidative insults, YB-1 assembly in SGs is associated with an enhancement of YB-1 protein secretion. An enriched fraction of extracellular YB-1 (exYB-1) significantly inhibited proliferation of receiving cells and such inhibition was associated to a G2/M cell cycle arrest, induction of p21WAF and reduction of ΔNp63α protein level. All together, these data show that acute oxidative stress causes sustained release of YB-1 as a paracrine/autocrine signal that stimulate cell cycle arrest.
Cell Communication and Signaling, Volume 16, pp 1-14; https://doi.org/10.1186/s12964-018-0274-6
Cold shock proteins are multifunctional RNA/DNA binding proteins, characterized by the presence of one or more cold shock domains. In humans, the best characterized members of this family are denoted Y-box binding proteins, such as Y-box binding protein-1 (YB-1). Biological activities range from the regulation of transcription, splicing and translation, to the orchestration of exosomal RNA content. Indeed, the secretion of YB-1 from cells via exosomes has opened the door to further potent activities. Evidence links a skewed cold shock protein expression pattern with cancer and inflammatory diseases. In this review the evidence for a causative involvement of cold shock proteins in disease development and progression is summarized. Furthermore, the potential application of cold shock proteins for diagnostics and as targets for therapy is elucidated.
International Journal of Molecular Sciences, Volume 19; https://doi.org/10.3390/ijms19061791
Non-melanoma skin cancers (NMSCs) are the leading cause of skin cancer-related morbidity and mortality. Effective strategies are needed to control NMSC occurrence and progression. Non-toxic, plant-derived extracts have been shown to exert multiple anti-cancer effects. Graviola (Annona muricata), a tropical fruit-bearing plant, has been used in traditional medicine against multiple human diseases including cancer. The current study investigated the effects of graviola leaf and stem extract (GLSE) and its solvent-extracted fractions on two human NMSC cell lines, UW-BCC1 and A431. GLSE was found to: (i) dose-dependently suppress UW-BCC1 and A431 cell growth, motility, wound closure, and clonogenicity; (ii) induce G0/G1 cell cycle arrest by downregulating cyclin/cdk factors while upregulating cdk inhibitors, and (iii) induce apoptosis as evidenced by cleavage of caspases-3, -8 and PARP. Further, GLSE suppressed levels of activated hedgehog (Hh) pathway components Smo, Gli 1/2, and Shh while inducing SuFu. GLSE also decreased the expression of pro-apoptotic protein Bax while decreasing the expression of the anti-apoptotic protein Bcl-2. We determined that these activities were concentrated in an acetogenin/alkaloid-rich dichloromethane subfraction of GLSE. Our data identify graviola extracts and their constituents as promising sources for new chemopreventive and therapeutic agent(s) to be further developed for the control of NMSCs.