Bladder cancer represents the most common malignancy of the urinary system, posing a significant threat to patients' life. Animal models and two-dimensional (2D) cell cultures, among other traditional models, have been used for years to study various aspects of bladder cancer. However, these methods are subject to various limitations when mimicking the tumor microenvironment in vivo, thus hindering the further improvement of bladder cancer treatments. Recently, three-dimensional (3D) culture models have attracted extensive attention since they overcome the shortcomings of their traditional counterparts. Most importantly, 3D culture models more accurately reproduce the tumor microenvironment in the human body because they can recapitulate the cell-cell and cell-extracellular matrix interactions. 3D culture models can thereby help us gain deeper insight into the bladder cancer. The 3D culture models of tumor cells can extend the culture duration and allow for co-culturing with different cell types. Study of patient-specific bladder cancer mutations and subtypes is made possible by the ability to preserve cells isolated from particular patients in 3D culture models. It will be feasible to develop customized treatments that target relevant signaling pathways or biomarkers. This article reviews the development, application, advantages, and limitations of traditional modeling systems and 3D culture models used in the study of bladder cancer and discusses the potential application of 3D culture models.
Endometritis is the inflammatory response of the uterine lining which is linked to infertility. Administration of platelet-rich plasma (PRP) represents a well-recommended strategy for the treatment of endometrium-associated infertility. In this study, we set to characterize the role and molecular mechanism of PRP intrauterine infusion in mice with endometritis.A mouse model of endometritis was established using lipopolysaccharide (LPS). Mouse endometrial epithelial cells were obtained in primary culture. PRP-treated cells were assayed for proliferative and apoptotic activities. Moreover, iNOS expression and chemokine and inflammatory factor contents in cells were assessed using RT-qPCR and ELISA. The mice were subjected to PRP intrauterine infusion. The expression of genes related to uterine development was analyzed by qPCR and the ki-67 content and caspase-3 activation in endometrial tissues were examined by immunohistochemistry. Finally, the Nrf2/HO-1 pathway activity in tissues was examined by Western blot.LPS induced inflammatory cell recruitment and tissue damage in the endometrium of mice, along with significantly increased levels of inflammatory and chemokine factors. PRP significantly enhanced endometrial epithelial cell activity, decreased apoptosis, and reduced inflammatory factor secretion. In addition, PRP intrauterine infusion significantly increased the expression of genes related to uterine development, promoted tissue proliferation, decreased apoptosis, and diminished inflammatory response in endometrial tissues of mice. PRP intrauterine infusion significantly elevated Nrf2/HO-1 pathway activity in endometrial epithelial cells and tissues.PRP intrauterine infusion significantly inhibited endometrial cell injury and alleviated the inflammatory response through activating the Nrf2/HO-1 pathway.
Antimicrobial resistance is a major global health problem that is developed upon exposure of bacteria to antimicrobial agents, and, thus, reducing or eliminating the ability of the currently available antibacterial drugs to eradicate bacterial infections. The aim of the current study was to encapsulate levofloxacin (third generation fluoroquinolones) into chitosan (CS) nanoparticles, to evaluate the antibacterial potency of the nanosized drug, and to characterize the major genetic mutations associated with the exposure of bacteria to the levofloxacin-loaded nanoparticle versus free levofloxacin. Three consecutive mutants were selected by stepwise exposure of one reference and two clinical Escherichia coli isolates to a series of progressively increasing concentrations of levofloxacin and the levofloxacin-loaded nanoparticles. Mutations in quinolone resistance determining region (QRDR) of gyrA and parC and regulators of AcrAB efflux pump (soxR, soxS and acrR) for all the selected-mutants were determined using polymerase chain reaction and sequencing. Mutants developed upon exposure to the nanosized drug had higher sensitivity to levofloxacin, compared with the levofloxacin-selected mutants. In addition, mutations in gyrA were observed in the levofloxacin first mutants, but in the nanosized levofloxacin second mutants. In the third mutants, an additional mutation in parC and mutations in the regulators were found only in levofloxacin-selected mutants. Loading of levofloxacin into the CS nanoparticles could increase the antibacterial activity of the drug and decrease the emergence of resistant mutants. To the best of our knowledge, this is the first study to explore the role of antimicrobial-loaded nanoparticles in the reduction of emergence of bacterial resistance.
We studied the associations and correlations between premature ejaculation (PE) and psychological disorders, such as anxiety and depression, in new perspectives with an aim of improving PE patients' treatment outcomes. Between December 2017 and December 2018, we selected 1,010 men aged over 18 years old. Self-estimated IELT, the premature ejaculation diagnostic tool, the International Index of Erectile Function-5, the General Anxiety Disorder-7 and the Patients Health Questionnaire-9 were used to measure latency time, premature ejaculation, erectile dysfunction, anxiety and depression respectively. Premature ejaculation patients were categorised into two types: lifelong PE (LPE) and acquired PE (APE). Among the 958 men evaluated, the prevalence of anxiety and depression in PE group was 82.07% (444/541) and 74.68% (404/541) respectively. Premature ejaculation patients after adjustment for age, negative association of IIEF-5 and positive relation of PEDT score with GAD-7/PHQ-9 were observed (p < 0.01 for all). These associations in men with LPE were stronger than APE. Stratification of the duration of PE showed that the longer the duration is, the more the prevalence of anxiety and depression will be. Age stratification showed that under the impact of PE, young men tend to have severe psychological problems.
Proliferative vitreoretinopathy (PVR) develops as a complication of rhegmatogenous retinal detachment (RRD). The unclear pathological pathways of PVR and RRD have been the biggest obstacle for successful retinal reattachment. In this study, a metabolomics approach using reversed-phase liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was developed to obtain a systematic view of the pathological processes of RRD and PVR. Through a partial least squares discriminant analysis (PLS-DA) method, 31 biomarkers were identified in the vitreous samples of RRD and PVR patients. Sixteen pathways participated in the development of RRD and PVR. Through analyzing the biological effects of those identified biomarkers, inflammation, proliferation and energy consumption were the three major disturbed biological processes involved in the RRD and PVR development. Inflammation happened in the pathological processes of RRD, and proliferation mainly happened during PVR formation. The established network of biomarkers indicated that the histidine metabolism and citrate cycle were seriously disturbed during the RRD and PVR development. The metabolomics study supplied a systematic view of the development and progression of RRD and PVR on a metabolite level.
Atropa belladonna is officially deemed as the commercial plant to produce scopolamine in China. In this study we report the simultaneous overexpression of two functional genes involved in biosynthesis of scopolamine, which encode the upstream key enzyme putrescine N-methyltransferase (PMT) and the downstream key enzyme hyoscyamine 6β-hydroxylase (H6H), respectively, in transgenic herbal plants Atropa belladonna. Analysis of gene expression profile indicated that both pmt and h6h were expressed at a higher level in transgenic lines, which would be favorable for biosynthesis of scopolamine. High-performance liquid chromatography result suggested that transgenic lines could produce higher accumulation of scopolamine at different levels compared with wild-type lines. Scopolamine content increased to 7.3-fold in transgenic line D9 compared with control lines. This study not only confirms that co-overexpression of pmt and h6h is an ideal method to improve the biosynthetic capacity of scopolamine but also successfully cultivates the transgenic line D9, which significantly enhanced the scopolamine accumulation. Our research can serve as an alternative choice to provide scopolamine resources for relative industry, which is more competitive than conventional market.
Anthocyanidin synthase (ANS) catalyzes the biosynthesis of anthocyanidin, which is a late gene for anthocyanin biosynthesis. In order to investigate the role of anthocyanidin synthase in anthocyanin biosynthesis, we cloned and characterized the anthocyanidin synthase gene from purple-flesh sweet potato (Ipomoea batatas (L.) Lam) Yuzi 263, which was designated as IbANS. The cDNA fragment of the ANS gene of sweet potato was 1375-bp in length which contained a 1086-bp open reading frame that encoded a 362-amino acid polypeptide. Comparative analysis showed that IbANS had a high similarity to other plant ANSs. The tissue expression profiles of IbANS indicated that it could be expressed in all tissues but at different levels. The higher expression level of IbANS was found in diameter (3.0 cm) of tuberous roots and periderms, while the lower expression level of IbANS was found in other tissues just coinciding with the anthocyanin content distribution.
The inwardly rectifying potassium channel protein Kir4.1 and the water channel protein aquaporin-4 (AQP4) have been suggested to play essential roles in the potassium and water homeostasis of the retina. In this study, we investigated the expression of Kir4.1 and AQP4 in the retina during endotoxin-induced uveitis (EIU) in rats.EIU was induced in male Wistar rats by intravitreal injection of lipopolysaccharide (LPS). The severity of the EIU was evaluated by clinical and histopathological examination. The expression of Kir4.1 and AQP4 in the retina was detected by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemical staining.In the animal model of EIU, the clinical changes correlated well with the histopathological findings. The inflammation peaked at 24 h and resolved by seven day. After an intravitreal LPS injection, the expression of Kir4.1 in the retina showed a significant decline at both the protein and mRNA levels. In the early stages of EIU, the expression of Kir4.1 mRNA decreased sharply, reaching a minimum at 12 h (31%, p<0.001). It then increased gradually and had partially recovered 14 days after LPS injection (92%, p>0.05). The expression of Kir4.1 protein decreased significantly, reaching a minimum at three days after the LPS injection (43%, p<0.001). Thereafter, it increased slightly but was maintained at a low level until 14 days after LPS injection (64%, p<0.001). In contrast, the expression of AQP4 mRNA remained almost unchanged after LPS treatment (p>0.05). The expression of AQP4 protein was only slightly reduced at one day (82%, p>0.05) after LPS injection and then increased gradually and had nearly recovered to the basal level at 14 days after LPS injection.EIU differently alters the expression of Kir4.1 and AQP4 in the retina. The differential expression of Kir4.1 and AQP4 during EIU implies a disturbance of water and potassium transport in the retina, which may contribute to the retinal edema during ocular inflammation.
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