Tunneling Proton Grotthuss Transfer Channels by Hydrophilic-Zincophobic Heterointerface Shielding for High-Performance Zn-MnO2 Batteries.

Hollandite-type manganese dioxide (α-MnO2) is recognized as a promising cathode material upon high-performance aqueous zinc-ion batteries (ZIBs) owing to the high theoretical capacities, high working potentials, unique Zn2+/H+ co-insertion chemistry, and environmental friendliness. However, its practical applications limited by Zn2+ accommodation, where the strong coulombic interaction and sluggish kinetics cause significant lattice deformation, fast capacity degradation, insufficient rate capability, and undesired interface degradation. It remains challenging to accurately modulate H+ intercalation while suppressing Zn2+ insertion for better lattice stability and electrochemical kinetics. Herein, proton Grotthuss transfer channels are first tunneled by shielding MnO2 with hydrophilic-zincophobic heterointerface, fulfilling the H+-dominating diffusion with the state-of-the-art ZIBs performance. Local atomic structure and theoretical simulation confirm that surface-engineered α-MnO2 affords to the synergy of Mn electron t2g-eg activation, oxygen vacancy enrichment, selective H+ Grotthuss transfer, and accelerated desolvation kinetics. Consequently, fortified α-MnO2 achieves prominent low current density cycle stability (≈100% capacity retention at 1 C after 400 cycles), remarkable long-lifespan cycling performance (98% capacity retention at 20 C after 12 000 cycles), and ultrafast rate performance (up to 30 C). The study exemplifies a new approach of heterointerface engineering for regulation of H+-dominating Grotthuss transfer and lattice stabilization in α-MnO2 toward reliable ZIBs.

Enhancing Phosphorus Release from Sewage Sludge in Anaerobic Digestion via Thermal Hydrolysis Pretreatment: Insights from Phosphorus Speciation and Molecular Biological Pathways.

This study explores the mechanisms enhancing phosphorus (P) release from sludge in anaerobic digestion (AD) with thermal hydrolysis pretreatment (THP) using sequential chemical extraction, X-ray absorption near-edge structure spectroscopy (XANES), 31P NMR, and multiomics. THP-treated sludge notably increased liquid-phase P by 53.8% over 3 days compared to sewage sludge (SS), identifying solid-phase Fe-P as the primary P source. The THP+AD also provided a higher abundance of bacteria that contributed to P release through multiple pathways (MPRPB), whereas SS+AD enriched some microbial species with single P release pathway. Moreover, species co-occurrence network analysis underlined the pivotal role of P-releasing bacteria in THP+AD, with 8 out of 16 keystones being P-releasers. Among the 63 screened genes that were related to P transformations and release, the poly beta-hydroxybutyrate (PHB) synthesis genes associated with polyphosphate bacteria-mediated P release were more abundant in THP+AD than in SS+AD. Furthermore, the upregulation of genes involved in methyl phosphonate metabolism in the THP-treated sludge enhanced the methane production potential of the AD process. These findings suggested that MPRPB were indeed the main contributors to P release, and enrichment in the THP+AD process enhanced their capability for P liberation.

The locoweed endophyte Alternaria oxytropis affects root development in Arabidopsis in vitro through auxin signaling and polar transport.

Locoweeds are leguminous forbs known for their toxicity to livestock caused by the endophytic fungi Alternaria sect. Undifilum. Unlike the defensive mutualisms reported in many toxin-producing endophytes and their plant hosts, the benefits that A. sect. Undifilum can confer to it host plants remains unclear. Here, we conducted physiological and genetic analyses to show that A. (sect. Undifilum) oxytropis influences growth, especially root development, in its locoweed host Oxytropis ochrocephala and Arabidopsis. The presence of A. oxytropis significantly decreased primary root length while increasing the numbers of lateral roots and root hairs, and increasing plant leaf area and fresh weight. The fungus also increased the concentrations of plant endogenous auxin, and the expression of key genes for auxin biosynthesis, signaling, and transport. These effects on root development were abolished in mutants deficient in auxin signaling and polar transport. Alternaria oxytropis down-regulated expression of PIN1 but increased expression of PIN2, PIN7, and AUX1, which might reflect alterations in the spatial accumulation of auxin responsible for the changes in root architecture. Plant growth was insensitive to A. oxytropis when naphthylphthalamic acid was applied. Our findings indicate a function of A. oxytropis in promoting the growth and development of Arabidopsis via the regulation of auxin, which in turn suggests a possible role in benefiting its locoweed hosts via a process independent of its toxin production.

Positive associations between different circulating trans fatty acids (TFAs) and urinary albumin excretion among adults in the U.S.: a population-based study.

BACKGROUND: It is well established that the consumption of trans-fatty acids (TFAs) can increase the incidence of total mortality, cardiovascular disease, cancer, and diabetes. However, there are still no demographic studies on the effects of circulating TFA isoforms on the albumin-creatinine ratio (ACR), an early marker of chronic kidney disease. Our goal was to explore the possible relationships between TFAs and ACR. METHODS: In this study, complete TFAs and urinary ACR data were collected from the National Health and Nutrition Examination Survey (NHANES) (2009-2010 and 1999-2000 cycles). The independent linear relationships between different circulating TFA isoforms and the ACR were examined by performing multivariable linear regression models. Machine learning was used to analyze the contribution of the different TFA isoforms to the ACR. To assess the nonlinearity of the relationship, smooth curve fitting and an analysis of threshold effect were performed, and a stratified analysis was conducted to identify possible susceptible populations. RESULTS: Our analysis included a total of 3785 individuals. Elaidic acid, linolelaidic acid, and sum TFAs were shown to be positively associated with the ACR after full adjustment by weighted multivariable regression analysis. In the subgroup analysis, the positive associations were maintained in participants with hypertension and without diabetes. In the XGBoost model of the ACR, Sum TFAs were found to be the most crucial factor. In addition, smooth curve fitting showed that there was a nonlinear relationship between the different TFAs and the ACR, and there was a saturation point. CONCLUSIONS: Our study demonstrated that TFA isoforms were positively and independently correlated with urinary albumin excretion, especially in participants with hypertension and without diabetes. This suggested that reducing trans fatty acid intake may reduce the risk of renal events.

Nutrient dynamics and microbial community response in macrophyte-dominated lakes: Implications for improved restoration strategies.

Although lakes dominated by macrophytes are conducive to ecological balance, this balance is easily disrupted by excessive nutrients flowing into the lake. However, knowledge of whether excessive nutrients lead to different microbial environmental vulnerabilities in the lake sediment between macrophyte-dominated areas and macrophyte-free areas is a prerequisite for the implementation of targeted protection measures. In this study, we investigated bacterial communities in sediments using high-throughput sequencing of 16S rRNA genes. Our results showed that the sources of total nitrogen (TN) and organic matter (OM) were related to the macrophytes. The structure, drivers, and interspecific associations of bacterial community, which were more susceptible to increased changes in TN and OM, differed significantly between macrophyte-dominated areas and macrophyte-free areas. More precisely, the lake edge, where was occupied by macrophytes, had a higher proportion of deterministic phylogenetic turnover (88.89%) than other sites, as well as a wider ecological niche and a tighter network structure. Further, as the difference in TN increased, the main assembly processes in surface sediments changed from stochastic to deterministic. However, the majority of phyla from the lake edge showed a greater correlation with excessive nutrients, and the selection of the community by excessive nutrients was more obvious at the edge of the lake. In addition, our results demonstrated that the stability of the bacterial community in macrophyte-free areas is greater than in macrophyte-dominated areas, while an excessively high deterministic process ratio and nutrient (TN and OM) concentration significantly reduced bacterial community stability at macrophyte-dominated areas. Taken together, these results provide a better understanding of the effects of excessive nutrients derived from macrophytes on bacterial community patterns, and highlight the importance of avoiding the accumulation of TN and OM in macrophyte-dominated areas to enhance the sustainability of the ecosystem after restoration of lakes with macrophytes.

Diet composition affects long-term zearalenone exposure on the gut-blood-liver axis metabolic dysfunction in mice.

Zearalenone (ZEN), one of the most contaminated Fusarium toxins worldwide, is very common in contaminating wheat, corn oil and other foods. People are more vulnerable to ZEN exposure with more daily caloric intake, yet little is known about the combined effect of different dietary patterns with mycotoxins. This study aimed to compare the effects of long-term ZEN exposure on the overall biochemical landscape of the "gut-blood-liver axis" under normal diet and high-fat diet (HFD) using a combined multi-omics approach. The results indicated that ZEN exposure, possibly via the phenylalanine metabolic pathway, led to dysbiosis of mouse flora, suppression of short-chain fatty acids (SCFAS) metabolism, systemic inflammatory responses, and disturbances in serum and liver metabolism, which were exacerbated in synergy with HFD and ultimately led to a more severe state of lipid metabolism in the liver. We further found that ZEN exposure attenuated the indole-3-propionic acid (IPA) metabolic pathway, enhanced 2-hydroxybutyric acid metabolism in serum, and attenuated ß-alanine metabolism in liver which was positively correlated with the abundance of Prevotellaceae UCG-004, Prevotellaceae UCG-001, and Prevotellaceae NK3B31 groups. The results highlighted the damaging effects of ZEN on the gut-blood-liver axis under different dietary patterns, which might serve as a reference for future studies exploring the combined effects of fungal toxins and multiple dietary patterns.

Simultaneous determination of multiple active 2-(2-phenylethyl)chromone analogues in agarwood by HPLC, QAMS, and UPLC-MS.

INTRODUCTION: Chromones are the major constituents of agarwood and are considered to be directly related to its quality. Agarotetrol, a chromone derivative, is a Chinese Pharmacopoeia content detection index. However, comprehensive high-performance liquid chromatography (HPLC), quantitative analysis of multiple components by a single marker (QAMS), and ultra-performance liquid chromatography mass spectrometry (UPLC-MS) analyses of this pharmacopeial plant material have never been performed. Moreover, reports regarding the separation and detection of multiple active 2-(2-phenylethyl)chromone analogues from this plant material are surprisingly scarce. OBJECTIVE: To establish a simple, reliable, and effective HPLC method utilising both diode array and MS detection for the simultaneous determination of multiple active chromone analogues in agarwood. METHODS: Four 2-(2-phenylethyl)chromones were isolated from methanol extracts of agarwood. After optimising the extraction, separation, and analytical conditions, validation of the developed analytical method indicated good linearity, satisfactory precision, and good recovery. On this basis, a method for the quantitative analysis of multiple components by a single marker was established. The four 2-(2-phenylethyl)chromones were identified by nuclear magnetic resonance spectroscopic analysis and UPLC coupled to electrospray ionisation quadrupole-time-of-flight MS. CONCLUSIONS: The behaviour of the chromones characterised by MS fragmentation indicated a loss of molecular CO and the formation of m/z 121 compounds by the cleavage of CH2 -CH2 bonds between the chromone and phenyl moieties. Three detection methods were successfully used in this study for agarwood detection, and this protocol may potentially be used as a tool for the quality control of agarwood.

Enhancement of the Aggregation-Induced Emission by Hydrogen Bond for Visualizing Hypochlorous Acid in an Inflammation Model and a Hepatocellular Carcinoma Model.

As an important reactive oxygen species, hypochlorous acid (HClO) is produced in various physiological processes. The abnormal rise of the HClO level is associated with a large number of inflammatory diseases. In this work, we develop a simple, aqueous-soluble aggregration-induced emission (AIE) probe for sensing HClO with significant aggregation-induced fluorescence (>1000 times). Two probes, CH3O-TPE-Py+-N+ (COTN) and OH-TPE-Py+-N+ (HOTN) (TPE, tetraphenylethylene), are synthesized for sensing HClO by the cleavage of the Py+-N+ group; the reaction products are CH3O-TPE-CHO (COT) and OH-TPE-CHO (HOT), respectively. The hydrophobicity of the probes is changed with the increased aggregation-induced emission. During the process, HOTN shows significantly better response than COTN. The slightly different chemical structures of COTN and HOTN result in a significant response to HClO. The theoretical calculation data support the theory that the hydrogen bond contributes to the excellent sensitivity for HClO. On the basis of the good response to HClO in vitro, HOTN is used to image inflammation and hepatocellular carcinoma in vivo because these diseases always produce high HClO levels.

Combined tetraphenylethylene fluorogens with positive charge for imaging capsule-covered pathogens.

Capsule-covered pathogens can cause serious infectious diseases, and are highly pathogenic to humans. Herein, we developed four positively charged tetraphenylethylene derivatives (PC-TPEgens) that in certain combinations were applied to identify capsule-bearing pathogens using fluorescence imaging. The dual-charged probes were used to visualize the entire process of phagocytosis of pathogens into macrophages.

New insights into impact of thermal hydrolysis pretreatment temperature and time on sewage sludge: Structure and composition of sewage sludge from sewage treatment plant.

This work investigated the effects of THP temperature (140-200 °C) and time (10-60 min) on the surface morphology, pyrolysis properties, and soluble compounds of dewatered sludge. Results indicate that higher temperature and longer pretreatment time considerably improve organics hydrolysis (the SCOD content increased 1.4-2.46 times, increasing 2.46 times at 200 °C). In addition, high temperature also improved the sludge pyrolysis efficiency (the highest at 200 °C), and reduced the harmful gas release, especially HCN. Moreover, the surface morphology of the sludge changed, the gap and floccules on the surface of the sludge increased. The carbohydrate content increased the highest; approximately 91.9% at 170 °C. THP promoted the decomposition of the nitrogen compounds in the sludge and facilitated their transition to a liquid phase. The total nitrogen and ammonia nitrogen content doubled, and the organic nitrogen content decreased by 50% with time increased from 10 to 60 min.

A Remotely Controlled Transformable Soft Robot Based on Engineered Cardiac Tissue Construct.

Many living organisms undergo conspicuous or abrupt changes in body structure, which is often accompanied by a behavioral change. Inspired by the natural metamorphosis, robotic systems can be designed as reconfigurable to be multifunctional. Here, a tissue-engineered transformable robot is developed, which can be remotely controlled to assume different mechanical structures for switching locomotive function. The soft robot is actuated by a muscular tail fin that emulates the swimming of whales and works as a cellular engine powered by the synchronized contraction of striated cardiac microtissue constructs. For a transition of locomotive behavior, the robot can be optically triggered to transform from a spread to a retracted form, which effectively changes the bending stiffness of the tail fins, thus minimizing the propulsion output from the "tail fin" and effectively switching off the engine. With the unprecedented controllability and responsiveness, the transformable robot is implemented to work as a cargo carrier for programmed delivery of chemotherapeutic agents to selectively eradicate cancer cells. It is believed that the realization of the transformable concept paves a pathway for potential development of intelligent biohybrid robotic systems.

HBO1 promotes cell proliferation in bladder cancer via activation of Wnt/ß-catenin signaling.

Histone acetyltransferase binding to ORC1 (HBO1), a histone acetyltransferase, was recently identified as an oncoprotein; however, its role in bladder cancer remains unknown. In this study, we showed that HBO1 was highly expressed at both the mRNA and the protein levels in bladder cancer. HBO1 expression was associated with the clinical features of human bladder cancer, including tumor size (P = 0.018) and T (P = 0.007) classifications. Patients with higher HBO1 expression had shorter recurrence-free survival time, whereas patients with lower HBO1 expression had better survival time. Moreover, we found that ectopic overexpression of HBO1 promoted, whereas HBO1 silencing inhibited tumor growth in bladder cancer cells both in vitro and in vivo. We further demonstrated that upregulation of HBO1 activated the Wnt/ß-catenin signaling pathway and led to nuclear localization of ß-catenin and upregulation of downstream targets of of Wnt/ß-catenin signaling. These findings suggest that HBO1 plays a key role in the progression of bladder cancer via the Wnt/ß-catenin pathway, and may serve as a potential therapeutic target for the treatment of bladder cancer.

Development of 12 microsatellite markers for Bombina orientails based on RNA-Seq and their usefulness in population genetic diversity.

Bombina orientalis is widely used due to bombesin which isolated from its skin. But in recent years, the population of B. orientalis has become declining distinctly because of human activities, environmental pollution, drought climatic conditions and other factors. In order to provide the molecular basis for the proposal of biodiversity conservation, we report the development of 12 microsatellite markers for B. orientalis based on RNA-Seq. We test polymorphism against in 48 B. orientalis individuals which randomly selected from 182 individuals take advantage of polyacrylamide gel electrophoresis (PAGE). These markers will be useful in the research on the genetic diversity, population genetic structure and other studies. For B. orientalis, all of these loci showed polymorphism, and in line with the H-W equilibrium law. The number of alleles per locus ranged from 3 to 21. The observed and expected heterozygosities ranged from 0.0118 to 0.7795 and from 0.1612 to 0.8703, respectively. The polymorphism information content ranged from 0.153 to 0.857. And the genetic diversity of B. orientalis in Lushui Rivers is significantly higher than that in the Maoer Mountains.

Two new sesquiterpenes from the aerial parts of Schizonepeta tenuifolia.

Two new sesquiterpenes named petafolias A-B were isolated from the aerial parts of Schizonepeta tenuifolia. Their structures were elucidated by various spectroscopic techniques (UV, IR, MS, CD, 1D, and 2D NMR).

Musashi-2 promotes migration and invasion in bladder cancer via activation of the JAK2/STAT3 pathway.

Musashi-2 (Msi2) is considered to have a crucial role in regulating various key cellular functions. However, the clinical significance and biological role of Msi2 in bladder cancer remains unknown. We examined the expression of Msi2 in bladder cancer cell lines in 167 clinical samples and the biological role of Msi2 in bladder cancer cells. Western blotting was used to investigate the possible mechanism of Msi2-induced migration and invasion in bladder cancer. Msi2 was significantly upregulated in bladder cancer cells and tissues compared with normal bladder urothelial cells and tissues. Immunohistochemical analysis revealed high expression of Msi2 in 57 of 167 (34.1%) bladder cancer specimens. Statistical analysis showed a significant correlation of Msi2 expression with advanced clinical stage, lymph node metastasis, and poor prognosis. Overexpression and ablation of Msi2 promoted and inhibited, respectively, the migration and invasion of bladder cancer cells. Furthermore, we found that Msi2 activated the JAK2/STAT3 pathway and promoted expression of genes downstream of JAK2/STAT3 in bladder cancer. This study demonstrates that Msi2 can induce bladder cancer cell migration and invasion by activating the JAK2/STAT3 pathway, and that Msi2 may be a valuable prognostic biomarker for bladder cancer patients.

Observations and analysis with the spline-based Rydberg-Klein-Rees approach for the 3(1)Σg(+) state of Rb2.

Ro-vibrational term values of the 3(1)Σg (+) state of (85,85)Rb2 and (85,87)Rb2 and resolved fluorescence spectra to the A(1)Σu (+) state are recorded following optical-optical double resonance excitation. The experimental data are heavily perturbed, and as a result, the standard analysis based on Dunham series representation of the energy levels fails. The analysis is done via modeling the adiabatic potential function with the Rydberg-Klein-Rees potential constructed from the generalized smoothing spline interpolation of the vibrational energies Gv and rotational constants Bv.

[Survey on fungi invasion of feed ingredients in 2013-2014 from parts of China].

OBJECTIVE: To survey the fungi contamination of four kinds of feed ingredients in 2013- 2014 from parts of China. METHODS: A total of 795 feed ingredients including soybean meal, cottonseed meal, wheat bran and distillers dried grains with soluble( DDGS) were collected from representative enterprises in different parts of China. Food safety national standards GB 4789. 15-2010 microbiological examination of food hygiene enumeration of molds and yeast and GB / T 4789. 16-2003 was used to enumerate, isolate and identify fungi, respectively. RESULTS: A total of 25 genus 54species kinds of fungi were isolated and the fungi from Aspergillus, Penicillium, Fusarium were the common contamination fungi in feed ingredients. Wheat bran was the most serious contaminated feed ingredients by fungi, and the detection rate of fungi contamination in four seasons were all greater than 84. 9%, and the exceeding feed limit rate was amount to 20. 8%. The fungi detection rate and contamination level were relatively lower in cottonseed meal and soybean meal, and the exceeding feed limit rate was 0. 9% and 1. 4%, respectively. There were types of feed ingredients, seasonal and regional differences for Aspergillus flavus and Fusarium moniliforme contamination. The detection rate of Aspergillus flavus in wheat bran was higher in four seasons and all surveyed areas, and autumn and winter and huazhong district were with the highest contamination level. The detection rate of Fusarium moniliforme for soybean meal in autumn and winter and huabei district was higher than others. The detection rate of Aspergillus flavus for DDGS was very low, but the detection rate of Fusarium moniliforme was higher, especially for autumn and huazhong district, the detection rate was 40. 4%and 50. 0%, respectively. CONCLUSION: The feed ingredients from China were commonly contaminated by fungi. It is recommended that strengthening the fungi contamination monitoring of feed ingredients from summer and autumn, huabei and huazhong district was an important prevention method.

Two NAC transcription factors from Caragana intermedia altered salt tolerance of the transgenic Arabidopsis.

BACKGROUND: Plants are continuously challenged by different environment stresses, and they vary widely in their adjustability. NAC (NAM, ATAF and CUC) transcription factors are known to be crucial in plants tolerance response to abiotic stresses, such as drought and salinity. ANAC019, ANAC055, and ANAC072, belong to the stress-NAC TFs, confer the Arabidopsis abiotic stress tolerance. RESULTS: Here we isolated two stress-responsive NACs, CiNAC3 and CiNAC4, from Caragana intermedia, which were induced by ABA and various abiotic stresses. Localization assays revealed that CiNAC3 and CiNAC4 localized in the nuclei, consistent with their roles as transcription factors. Histochemistry assay using Pro(CiNAC4)::GUS transgenic Arabidopsis showed that the expression of the GUS reporter was observed in many tissues of the transgenic plants, especially in the root vascular system. Overexpression of CiNAC3 and CiNAC4 reduced ABA sensitivity during seed germination, and enhanced salt tolerance of the transgenic Arabidopsis. CONCLUSIONS: We characterised CiNAC3 and CiNAC4 and found that they were induced by numerous abiotic stresses and ABA. GUS histochemical assay of CiNAC4 promoter suggested that root, flower and local damaged tissues were the strongest stained tissues. Overexpression assay revealed that CiNAC4 play essential roles not only in promoting lateral roots formation, but also in responding to salinity and ABA treatment of Arabidopsis.

The sesquiterpene biosynthesis and vessel-occlusion formation in stems of Aquilaria sinensis (Lour.) Gilg trees induced by wounding treatments without variation of microbial communities.

As widely recognized, agarwood formation in Aquilaria trees is induced by external wounding. Because agarwood usually harbors specific microbes, the function of microbes in agarwood formation has been debated for almost a century. In this study, two wounding methods, the burning-chisel-drilling method (BCD) and the whole-tree agarwood-inducing method (Agar-Wit), were used under the non-contamination of environmental microorganisms. After pyrosequencing the small rRNA subunits of the wounds induced by the BCD and Agar-Wit, no substantial variation was observed either in fungal and bacterial enrichment and diversity or in the relative abundances of taxa. By contrast, significant variations in fungal and bacterial communities were detected following the partial tree pruning (PTP)-wounding. The wound-induced sesquiterpene biosynthesis and vessel-occlusion formation, however, were found to be similar in all types of wounded trunks. We thus infer that wounding in the absence of variations in microbial communities may induce agarwood formation. This result does not support the long-standing notion that agarwood formation depends on microbes.

Long-term versus short-term introvesical chemotherapy in patients with non-muscle-invasive bladder cancer: a systematic review and meta-analysis of the published results of randomized clinical trials.

In order to assess the effect of long-term versus short-term intravesical chemotherapy in preventing the recurrence of patients with non-muscle-invasive bladder cancer, we searched several databases with words as mesh terms and free text words to find all eligible randomized clinical trials (RCTs) for the comparison of the two strategies of instillation durations. "Observed-Expected events research (O-E)" and "Variance (V)" for calculating hazard ratio (HR) were used in Revman 5.2 software recommended by Cochrane Collabration for data analysis. Sensitivity and subgroup analysis were selected to minish heterogeneity. GRADEpro 3.6 profile recommended by Cochrane Collabration was employed for quality assessment of analyses. Finally, 13 eligible RCTs with 4216 patients were included in this review and 16 comparisons from 13 trials were involved for analysis. The pooled analysis revealed no significant difference between long-term and short-term duration [HR=0.99, 95% CI (0.89, 1.11), P=0.89]. Within the subgroup analysis, patients benefited from long-term instillations with a start regimen of one immediate instillation [HR=0.83, 95% CI (0.69, 1.00), P=0.05]. But patients were not suitable to receive long-term instillations with epirubicin (EPI) [HR=1.01, 95% CI (0.91, 1.13), P=0.78]. The progression rate was not reduced after long-term instillations [HR=0.96, 95% CI (0.66, 1.39), P=0.82]. From our results, patients should not receive introvesical chemotherapy more than half a year. In contrast, patients with one immediate instillation are preferred to have a long-term duration at least one year. Long-term instillations can not reduce the progression rate.