Use of municipal solid waste incineration fly ash as a supplementary cementitious material: CO2 mineralization coupled with mechanochemical pretreatment.

The use of municipal solid waste incineration fly ash, commonly referred to as "fly ash", as a supplementary cementitious material (SCM), has been explored to mitigate the CO2 emissions resulting from cement production. Nevertheless, the incorporation of fly ash as an SCM in mortar has been shown to weaken its compressive strength and increase the risk of heavy metal leaching. In light of these challenges, this study aims to comprehensively evaluate the influence of CO2 pressure, temperature, and residual water/binder ratio on the CO2 uptake and compressive strength of mortar when combined with fly ash. Additionally, this study systematically examines the feasibility of mechanochemical pretreatment, which enhances the homogenization of fly ash and augments the density of the mortar's microstructure. The results indicate that the use of mechanochemical pretreatment leads to a notable 43.6% increase in 28-day compressive strength and diminishes the leaching of As, Ba, Ni, Pb, Se, and Zn by 17.9-77.8%. Finally, a reaction kinetics model is proposed to elucidate the CO2 sequestration process under varying conditions. These findings offer valuable guidance for incorporating fly ash as an SCM and CO2 sequestrator in mortar.

SNHG1, a KLF4-upregulated gene, promotes glioma cell survival and tumorigenesis under endoplasmic reticulum stress by upregulating BIRC3 expression.

Increasing evidence indicates that long noncoding RNAs (lncRNAs) play crucial roles in the resistance to endoplasmic reticulum (ER) stress in many cancers. However, ER stress-regulated lncRNAs are still unknown in glioma. In the present study, we investigated the altered lncRNAs upon ER stress in glioma and found that small nucleolar RNA host gene 1 (SNHG1) was markedly increased in response to ER stress. Increased SNHG1 suppressed ER stress-induced apoptosis and promoted tumorigenesis in vitro and in vivo. Further mechanistic studies indicated that SNHG1 elevated BIRC3 mRNA stability and enhanced BIRC3 expression. We also found that KLF4 transcriptionally upregulated SNHG1 expression and contributed to the ER stress-induced SNHG1 increase. Collectively, the present findings indicated that SNHG1 is a KLF4-regulated lncRNA that suppresses ER stress-induced apoptosis and facilitates gliomagenesis by elevating BIRC3 expression.

Peptidomic analysis on synovial tissue reveals galectin-1 derived peptide as a potential bioactive molecule against rheumatoid arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease that leads to small joints irreversible destruction. Despite intense efforts, the pathophysiology of RA currently remains unclear. We aimed to gain insight into the pathophysiology process in peptidomic perspective and to identify bioactive peptides for RA treatment. METHODS: The endogenous peptides in synovial tissue between control and rheumatoid arthritis group were identified by liquid chromatography-mass spectrometry (LC-MS/MS). Since the biological function of peptides were always associated with precursor proteins, the potential function of the differentially peptides were predicted by GO and pathway analysis of their precursors. Besides, peptides located in the domains of their precursors were identified. Finally, we determined the impact of galectin-1 derived peptide by administration on the damage to MH7A cells caused by TNF-α. RESULTS: Totally, 141 down-regulated peptides and 10 up-regulated peptides were identified (Fold change > 1.5 and P < 0.05). It indicated that these differentially peptides were tightly involved in the pathophysiology process of RA preliminarily. Finally, we identified a peptide derived from the domain of galectin-1 could inhibit the abnormal proliferation induced by TNF-α and promoted apoptosis of MH7A. CONCLUSION: In summary, our study provided a better understanding of endogenous peptides in RA. We found a peptide that might be used in anti-RA treatment.

Bazedoxifene protects cerebral autoregulation after traumatic brain injury and attenuates impairments in blood-brain barrier damage: involvement of anti-inflammatory pathways by blocking MAPK signaling.

OBJECTIVE: Traumatic brain injury (TBI) is a significant cause of death and long-term deficits in motor and cognitive functions for which there are currently no effective chemotherapeutic drugs. Bazedoxifene (BZA) is a third-generation selective estrogen receptor modulator (SERM) and has been investigated as a treatment for postmenopausal osteoporosis. It is generally safe and well tolerated, with favorable endometrial and breast safety profiles. Recent findings have shown that SERMs may have therapeutic benefits; however, the role of BZA in the treatment of TBI and its molecular and cellular mechanisms remain poorly understood. The aim of the present study was to examine the neuroprotective effects of BZA on early TBI in rats and to explore the underlying mechanisms of these effects. MATERIALS AND METHODS: TBI was induced using a modified weight-drop method. Neurological deficits were evaluated according to the neurological severity score (NSS). Morris water maze and open-field behavioral tests were used to test cognitive functions. Brain edema was measured by brain water content, and impairments in the blood-brain barrier (BBB) were evaluated by expression analysis of tight junction-associated proteins, such as occludin and zonula occludens-1 (ZO-1). Neuronal injury was assessed by hematoxylin and eosin (H&E) staining. LC-MS/MS analysis was performed to determine the ability of BZA to cross the BBB. RESULTS: Our results indicated that BZA attenuated the impaired cognitive functions and the increased BBB permeability of rats subjected to TBI through activation of inflammatory cascades. In vivo experiments further revealed that BZA provided this neuroprotection by suppressing TBI-induced activation of the MAPK/NF-κB signaling pathway. Thus, mechanically, the anti-inflammatory effects of BZA in TBI may be partially mediated by blocking the MAPK signaling pathway. CONCLUSIONS: These findings suggest that BZA might attenuate neurological deficits and BBB damage to protect against TBI by blocking the MAPK/NF-κB signaling pathway.

The sodium pump α1 subunit regulates bufalin sensitivity of human glioblastoma cells through the p53 signaling pathway.

Bufalin is the primary component of the traditional Chinese medicine "Chan Su," which has been widely used for cancer treatment at oncology clinics in certain countries. Evidence suggests that this compound possesses potent antitumor activities, although the exact molecular mechanism(s) require further elucidation. Therefore, this study aimed to further clarify the in vitro and in vivo antiglioma effects of bufalin and the molecular mechanism underlying the regulation of drug sensitivity. The anticancer effects of bufalin were determined by colony formation assays, apoptosis assays, and cellular redox state tests of glioma cells. Confocal microscopy was performed to determine the expression changes of the DNA damage biomarker γ-H2AX and the nuclear translocation of p53 in glioma cells. Western blotting and RT-PCR were used to detect the protein and gene expression levels, respectively. Here, we report that bufalin induced glioblastoma cell apoptosis and oxidative stress and triggered DNA damage. The critical roles of the sodium pump α1 subunit (ATP1A1) in mediating the XPO1-targeted anticancer effect of bufalin in human glioma were further confirmed. Mechanistic studies confirmed the important roles of Src and p53 signaling in mediating bufalin-induced apoptosis. Importantly, bufalin also inhibited the growth of glioma xenografts. In conclusion, our study indicated that therapies targeting the ATP1A1 and p53 signaling-mediated mitochondrial apoptotic pathways regulated by bufalin might be potential treatments for human glioma, and these findings will provide molecular bases for developing bufalin into a drug candidate for the treatment of malignant glioma.

Anti-parasitic effect on Toxoplasma gondii induced by a spider peptide lycosin-I.

Toxoplasmosis is a widely distributed parasitic protozoan disease, caused by Toxoplasma gondii (T. gondii). High prevalence of toxoplasmosis and limitations of conventional treatments lead to a search for new therapeutic drugs. Lycosin-I is a linear peptide, derived from the venom of the spider Lycosa singoriensis. The aim of the present study was to determine the anti-parasitic effect of lycosin-Ι against T. gondii. In vitro, the anti-T. gondii activities of lycosin-Ι were evaluated by MTT assay, trypan blue exclusion assay, cell counting assay and plaque assay. Cytokines of IL-6 and IL-8 were measured by quantitative PCR. In addition, the structures of tachyzoites treated with lycosin-Ι were also observed by scanning and transmission electron microscopy. In vivo, mice were challenged with parasites treated by lycosin-I. The results revealed that lycosin-Ι had shown a significant ability to inhibit T. gondii invasion and proliferation. Cytokines of IL-6 and IL-8 were reduced by lycosin-Ι at transcription level in human foreskin fibroblast (HFF) cells infected with T. gondii tachyzoites, but they were increased compared to non-infected cells. For tachyzoites, lycosin-Ι induced their cell membrane alterations with formation of invaginations, some of them appeared to be vacuolated in their cytoplasm. Moreover, lycosin-Ι had prolonged the survival time of mice by controlling T. gondii proliferation. In conclusion, our present study provides the first evidence for anti-T. gondii by using the spider peptide lycosin-Ι. These findings suggest that lycosin-Ι is a potential alternative agent for the treatment of toxoplasmosis.

Fibular strut allograft influences reduction and outcomes after locking plate fixation of comminuted proximal humeral fractures in elderly patients: a retrospective study.

BACKGROUND: Proximal humeral fractures (PHFs) are the third most commonly occurring fractures in elderly patients. Most of these fractures can be treated with conservative methods, but the optimal surgical treatment strategy for unstable fractures in elderly patients remains controversial. This study aimed to compare the radiological and clinical outcomes between locking compression plate (LCP) fixation and LCP fixation with fibular allograft implantation for the treatment of comminuted PHFs. METHODS: We retrospectively reviewed 60 patients (mean age, 72.75 years) with closed 3- or 4-part fractures, and a minimum of 2 years of follow-up. Fracture reduction was quantitatively determined by humeral head height (HHH) and neck-shaft angle (NSA). The clinical outcome was evaluated by Constant-Murley score (CMS) and American Shoulder and Elbow Surgeons (ASES) score. RESULT: The average radiological changes were higher in the LCP group than in the locking plate with fibular allograft group (HHH of 4.16 mm vs 1.18 mm [p < 0.001] and NSA of 9.94° versus 3.12° [p < 0.001]) . The final average outcome scores were lower in the LCP group than in the FA group (CMS of 73.00 vs 78.96 [p = 0.024] and ASES score of 72.80 vs 78.64 [p = 0.022]). The FA group showed better forward elevation (p = 0.010) and abduction (p = 0.002); however, no significant differences were observed for shoulder external rotation or internal rotation. The number of complications was higher in the LCP group (28.57%) than in the FA group (1.2%) (p < 0.001). CONCLUSION: For comminuted PHFs in elderly patients, LCP fixation combined with a fibular allograft is reasonable option to ensure satisfactory radiological and clinical outcomes. TRIAL REGISTRATION: ZDYJLY(2018)New-9 . Name of registry: IEC for clinical Research of Zhongda Hospital, Affiliated to Southeast University. Date of registration: 2018-05-17.

Long non-coding RNA AC023115.3 suppresses chemoresistance of glioblastoma by reducing autophagy.

Malignant glioma is an aggressive brain cancer that responds poorly to chemotherapy. However, the molecular mechanism underlying the development of chemoresistance in glioma is not well-understood. In this study, we show that long non-coding RNA AC023115.3 is induced by cisplatin in human glioblastoma cells and that elevated AC023115.3 promotes cisplatin-induced apoptosis by inhibiting autophagy. Further mechanistic studies revealed that AC023115.3 acts as a competing endogenous RNA for miR-26a and attenuates the inhibitory effect of miR-26a on GSK3ß, a proline-directed serine-threonine kinase that promotes the degradation of Mcl1, leading to an increase in GSK3ß and a decrease in autophagy. Additionally, we discovered that AC023115.3 improves chemosensitivity of glioma cells to cisplatin by regulating the miR-26a-GSK3ß-Mcl1 pathway. Thus, these data indicate that the AC023115.3-miR-26a-GSK3ß signalling axis plays an important role in reducing the chemoresistance of glioma.

LINC01510 suppresses cell proliferation and invasion by inhibiting Wnt/ß-catenin signaling in renal cell carcinoma.

The role of long non-coding RNA in Renal cell carcinoma (RCC) tumorigenesis and progression remains largely unknown. Here, we found that LINC01510 functions as a tumor suppressor in RCC tumorigenesis. We screened TCGA database and then found that LINC01510 is significantly down-regulated in malignant RCC tissues, and the lower expression of LINC01510 predicts poor prognosis. Moreover, the down-regulated LINC01510 was further confirmed in our fresh tissues and cell lines. Biological functions assays shown that Ectopic expression of LINC01510 not only inhibits RCC cell proliferation both in vitro and in vivo, but also impairs cell invasion ability. Moreover, we found overexpression of LINC01510 inhibits the expression of CCND1 and CCNE1, as well as MMPs (MMP2, MMP7 and MMP9), and thus affecting RCC cell cycle and invasion. Meanwhile, Western blot assays revealed that the expression of ß-catenin is regulated by LINC01510; overexpression of ß-catenin could partly rescue the cell viability and invasion ability caused by ectopic expression of LINC01510. Taken together, we found that LINC01510 regulates cell proliferation and invasion by modulating Wnt/ß-catenin signaling in RCC.

Update on the effect of exogenous hormone use on glioma risk in women: a meta-analysis of case-control and cohort studies.

Various studies have confirmed the important roles of endogenous hormones in the development of gliomas, while the roles of exogenous hormones remain controversial. Based on case-control studies and cohort studies, a meta-analysis was exerted to explore the effect of two exogenous hormones use (HRT: hormone replacement therapy; OC: oral contraceptives) on glioma risk. 16 eligible studies, including 11 case-control studies and 5 cohort studies, containing 8055027 women, were included in our study. All included studies have reported the relative risks (RRs) or odds ratios (ORs), and 95% confidence intervals (CIs). We use the fixed-effects model to calculate the estimated overall risk. In case-control studies, the risk of glioma was lower in women who had ever been treated with an exogenous hormone than in the control group (HRT: OR 0.91, 95% CI 0.84-0.99; OC: OR 0.99, 95% CI 0.91-1.07). In research of cohort studies, similar results have been obtained (HRT: RR 0.95, 95% CI 0.83-1.08; OC: RR 0.75, 95% CI 0.66-0.84). Our study further confirmed that the use of exogenous hormones has an important impact on the risk of glioma in women. However, more prospective studies are needed to further confirm this conclusion.

The peptide lycosin-I attenuates TNF-α-induced inflammation in human umbilical vein endothelial cells via IκB/NF-κB signaling pathway.

OBJECTIVE: The peptide lycosin-I has anti-bacterial and anti-cancer capacities. However, the anti-inflammatory activity of lycosin-I remains unknown. We investigated whether lycosin-I could attenuate inflammation. MATERIALS AND METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with lycosin-I before exposure to tumor necrosis factor-α (TNF-α). The expression of intercellular cell adhesion molecule-1 (ICAM-1), nuclear transcription factor-kappa B (NF-κB) p65 and inhibitory subunit of NF-κB alpha (IκBα) was evaluated by western blot. The expression of interleukin-6 (IL-6) and interleukin-8 (IL-8) was detected by quantitative RT-PCR or ELISA. Immunofluorescence analysis was used to determine the impact of lycosin-I on NF-κB pathway. C57BL/6 mice were pretreated with lycosin-I before exposure with lipopolysaccharide (LPS). RESULTS: Lycosin-I significantly reduced the TNF-α-enhanced expression of IL-6, IL-8 and ICAM-1. Lycosin-I also inhibited the human monocyte cells adhesion to HUVECs. We further demonstrated that lycosin-I could effectively suppress the reaction of endothelial cells to TNF-α by inhibiting IκBα degradation. Subsequently, the phosphorylation and translocation of NF-κB p65 could also be attenuated. Furthermore, lycosin-I exhibited a significant protection of C57BL/6 mice against LPS-induced death. CONCLUSIONS: Our results suggested that the anti-inflammatory activity of lycosin-I was associated with NF-κB activation and lycosin-I had potential to be a novel therapeutic candidate for inflammatory diseases.

Improvement of pregnancy outcome by extending embryo culture in IVF-ET during clinical application.

PURPOSE: The purpose of this study is to investigate the application value of the extended embryo culture for 7-8 h in day 3 morning during IVF-ET process. METHODS: Embryos were retrospectively assessed during 08:00-09:00 on the morning of day 3 in the control group, and were assessed once again at 16:00 in the afternoon in the extended culture (EC) group. The embryos with good developmental potential were preferentially selected to transfer. The cumulative pregnancy outcomes were analyzed in one oocyte retrieval cycle. RESULTS: Similar proportions were found in the rates of cumulative clinical pregnancy, cumulative live birth, and the perinatal/neonatal outcomes per oocyte retrieval cycle (P > 0.05). But higher total clinical pregnancy rate, higher total implantation rate, and lower total abortion rate were obtained in the EC group (P < 0.05). After EC, 53.58% of the embryos were able to continue to develop. The transferred embryos were mainly composed of ≥ 8-cell embryos (75.90%) in the EC group and ≤ 8-cell embryos (82.92%) in the control group. Interestingly, the implantation rates were increasingly improved with the increasing blastomere number up to 56.31% at the morula stage in the EC group, while they were limited to 32.33% at 8-cell stage in the control group. CONCLUSIONS: The extended culture of day 3 embryos for 7-8 h not only reduced the risk of IVF-ET treatment compared to blastocyst culture through another 2-3 days, but also improved the clinical outcomes and the efficiency of every transferred cycle and every transferred embryo.

Correlation of VEGF and EGFR in peripheral blood with clinical stage and pathological grade of renal cell carcinoma and analysis of prognosis.

PURPOSE: To detect the expression of VEGF and EGFR in peripheral blood and cancer tissues of patients with renal cell carcinoma (RCC), and to explore the correlations with clinical stage, pathological grade and prognosis of disease. METHODS: A total of 64 patients with RCC who were diagnosed and treated from June 2016 to August 2017 in our hospital were enrolled. Patients were divided into different clinical stages and pathological grades, and ELISA and immunohistochemistry were used to detect the expression of VEGF and EGFR in peripheral blood. Peripheral blood was also taken from 24 healthy individuals to serve as control group. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of VEGF and EGFR in RCC tissues and paracancer tissues. All patients were followed up after discharge to record their survival. RESULTS: Significant differences in the expression levels of VEGF and EGFR were found between stage III and IV (p<0.05), but not between stage I and II. Expressions level of VEGF and EGFR in serum of well-differentiated, moderatelydifferentiated, and poorly-differentiated RCC were all higher than those in the healthy control group, and significant differences were found between different pathological grades (p<0.05). Patients with higher expression levels of VEGF and EGFR showed shorter survival compared to patients with lower expression levels (p<0.05). CONCLUSION: VEGF and EGFR in peripheral blood can be used as one of the effective indicators of prognosis of RCC. Our study provided reference for clinical treatment and prediction of prognosis of RCC.

Sulfiredoxin May Promote Cervical Cancer Metastasis via Wnt/ß-Catenin Signaling Pathway.

The abnormal elevation of sulfiredoxin (Srx/SRXN1)-an antioxidant enzyme whose main function is to protect against oxidative stress-has been shown to be closely correlated with the progression of several types of cancer, including human cervical cancer. However, the molecular mechanism by which Srx promotes tumor progression, especially cancer metastasis in cervical cancer, has not been elucidated. Here, we show that Srx expression gradually increases during the progression of human cervical cancer and its expression level is closely correlated with lymph node metastasis. Our study also reveals a significant positive correlation between the expression of Srx and ß-catenin in cervical cancer tissues. Loss-of-function studies demonstrate that Srx knockdown using a lentiviral vector-mediated specific shRNA decreases the migration and invasion capacity in HeLa (human papilloma virus 18 type cervical cancer cell line) and SiHa SiHa (cervical squamous cancer cell line). Notably, the exact opposite effects were observed in gain-of-function experiments in C-33A cells. Mechanistically, downregulation or upregulation of Srx leads to an altered expression of proteins associated with the Wnt/ß-catenin signaling pathway. Furthermore, blockage of the Wnt/ß-catenin signaling pathway contributed to attenuated Srx expression and resulted in significant inhibition of cell migration and invasion in cervical cancer cell lines. Combined, Srx might be an oncoprotein in cervical cancer, playing critical roles in activating the Wnt/ß-catenin signaling pathway; it may therefore be a therapeutic target for cervical cancer.

Is the digital economy empowering high-quality tourism development? A theoretical and empirical research from China.

How digital economy (DE) empowers high-quality development of tourism (HQDT) has become a common concern among scholars. Given this, this study clarifies the theoretical connotation of DE enabling HQDT,and finds that: Micro, DE promotes efficiency improvements in tourism enterprises, with its economies of scale and Matthew effect reducing average costs, its economies of scope meeting diversified demand, and its long-tail effect improving supply-demand matching mechanism. Meso, DE can transform and upgrade tourism industry structure through industrial digitization and digital industrialization, and also form a new tourist industry form and value chain through cross-border integration. Macro, DE can stimulate innovation and flexibility of market players, increase new factor inputs in tourism, improve factor allocation efficiency, and advance macro regulation of the tourism market. Accordingly, the study conducts an empirical test based on panel data for 31 provinces in mainland China during 2011-2020. Results show that: ① DE positively influences HQDT, and the sub-dimensions all positively influence HQDT. ② DE has a heterogeneous impact on HQDT and shows spatial spillover effects. Finally, the study concludes with effective paths for DE promoting HQDT: "Promote digital infrastructure construction, accelerate tourism digital transformation, strengthen integration and innovation development, and overcome the challenges of tourism enterprises".

Advancing Organoid Engineering for Tissue Regeneration and Biofunctional Reconstruction.

Tissue damage and functional abnormalities in organs have become a considerable clinical challenge. Organoids are often applied as disease models and in drug discovery and screening. Indeed, several studies have shown that organoids are an important strategy for achieving tissue repair and biofunction reconstruction. In contrast to established stem cell therapies, organoids have high clinical relevance. However, conventional approaches have limited the application of organoids in clinical regenerative medicine. Engineered organoids might have the capacity to overcome these challenges. Bioengineering-a multidisciplinary field that applies engineering principles to biomedicine-has bridged the gap between engineering and medicine to promote human health. More specifically, bioengineering principles have been applied to organoids to accelerate their clinical translation. In this review, beginning with the basic concepts of organoids, we describe strategies for cultivating engineered organoids and discuss the multiple engineering modes to create conditions for breakthroughs in organoid research. Subsequently, studies on the application of engineered organoids in biofunction reconstruction and tissue repair are presented. Finally, we highlight the limitations and challenges hindering the utilization of engineered organoids in clinical applications. Future research will focus on cultivating engineered organoids using advanced bioengineering tools for personalized tissue repair and biofunction reconstruction.

Ferroelectric switchable behavior through fast reversible de/adsorption of water spirals in a chiral 3D metal-organic framework.

A polar homochiral 3D MOF [{Co2(L)(bpe)(H2O)}·5H2O]n constructed with cobalt(II) and a new ligand N-(1,3-dicarboxy-5-benzyl)-carboxymethylglycine (H4L) accommodates ordered helical water streams in its helical grooves. It provides the first example of switchable ferroelectric and optical behavior through two-step reversible single-crystal to single-crystal transformation (SCSC) upon desorption/adsorption of water spirals and coordinated water molecules, respectively.

Asymmetrical/symmetrical D-π-A/D-π-D thiazole-containing aromatic heterocyclic fluorescent compounds having the same triphenylamino chromophores.

A family of linear asymmetrical D-π-A and symmetrical D-π-D types of thiazole-based aromatic heterocyclic fluorescent compounds bearing various electron-donating and electron-withdrawing tails (bromo, triphenylamino, pyridyl, thienyl and benzoic acid) have been designed and prepared successfully. Synthetic, structural, thermal, spectral and computational comparisons have been carried out for related compounds because of their adjustable electronic properties. It is interesting to mention that compound 2 can be prepared from 5-bromothiazole by one-pot Suzuki-Miyaura coupling and subsequent C-H activation reactions via a 5-TPA-substituted thiazole intermediate 1. X-ray single-crystal structures of six compounds indicate that they all crystallize in the triclinic P1 space group and the thiazole core exhibits different dihedral angles with its adjacent benzene ring of the triphenylamino group (3.6(3)-40.8(3)°). The photophysical and electrochemical results demonstrate that compound 7 exhibits high electrochemical activity with a green fluorescence emission. Meanwhile, compounds 1, 2, and 6 show high luminescence quantum yields, and compound 8 exhibits excellent thermal stability (T(d(10)) = 503 °C).

Development of a native-locus dual reporter system for the efficient screening of the hyper-production of natural products in Streptomyces.

Streptomyces is renowned for its abundant production of bioactive secondary metabolites, but most of these natural products are produced in low yields. Traditional rational network refactoring is highly dependent on the comprehensive understanding of regulatory mechanisms and multiple manipulations of genome editing. Though random mutagenesis is fairly straightforward, it lacks a general and effective strategy for high throughput screening of the desired strains. Here in an antibiotic daptomycin producer S. roseosporus, we developed a dual-reporter system at the native locus of the daptomycin gene cluster. After elimination of three enzymes that potentially produce pigments by genome editing, a gene idgS encoding the indigoidine synthetase and a kanamycin resistant gene neo were integrated before and after the non-ribosomal peptidyl synthetase genes for daptomycin biosynthesis, respectively. After condition optimization of UV-induced mutagenesis, strains with hyper-resistance to kanamycin along with over-production of indigoidine were efficiently obtained after one round of mutagenesis and target screening based on the dual selection of the reporter system. Four mutant strains showed increased production of daptomycin from 1.4 to 6.4 folds, and significantly improved expression of the gene cluster. Our native-locus dual reporter system is efficient for targeting screening after random mutagenesis and would be widely applicable for the effective engineering of Streptomyces species and hyper-production of these invaluable natural products for pharmaceutical development.

Development and validation of a novel nomogram model for predicting the survival of patients with T2-4a, N0-x, M0 bladder cancer: a retrospective cohort study.

OBJECTIVE: Recent developments in bladder cancer treatment strategies have significantly improved the prognosis of clinically curable muscle invasive bladder cancer (MIBC) patients. Here, the prognostic factors of T2-4a, N0-x, M0 MIBC patients were investigated using the Surveillance, Epidemiology, and End Results (SEER) database and a novel nomogram model was established for prognosis prediction. METHODS: The data of 7,292 patients with T2-4a, N0-x, M0 MIBC were retrieved from the SEER database (2000-2020) and randomly classified into a training set (n = 5,106) and validation set (n = 2,188). Kaplan-Meier analysis was used to calculate cancer-specific survival (CSS) and overall survival (OS) rates of patients, and differences between survival curves were analyzed using the log-rank test. Cox regression analysis was used to screen and incorporate patient prognosis-affecting independent risk factors into the nomogram model. Consistency index (C-index) values and areas under the time-dependent receiver operating characteristic curve (AUC) were used to evaluate the discriminatory ability, and the calibration curve was used to assess the calibration of the model. Its predictive performance and American Joint Committee on Cancer (AJCC) stage were compared using decision curve analysis (DCA). RESULTS: The 1-, 3-, and 5-year CSS and OS rates of patients with T2-4a, N0-x, M0 MIBC were 76.9%, 56.0%, and 49.9%, respectively, and 71.3%, 47.9%, and 39.5%, respectively. Cox regression analysis showed that age, marital status, race, pathological type, tumor size, AJCC stage, T stage, N stage, surgery of primary tumor, regional lymph node dissection, radiation, and chemotherapy were independent prognostic risk factors of both CSS and OS (P < 0.05). The C-index and AUC of the nomogram model constructed based on the training and validation sets were both > 0.7, and calibration curves for predicting the 1-, 3-, and 5-year survival were consistent with the ideal curve. The nomogram model showed a higher net benefit with DCA than AJCC stage analysis. CONCLUSION: The nomogram model could accurately predict the prognosis of patients with T2-4a, N0-x, M0 MIBC. It may help clinicians perform personalized prognosis evaluations and formulate treatment plans.