Tolerance threshold of a pelagic species in China to total dissolved gas supersaturation: from the perspective of survival characteristics and swimming ability.

Total dissolved gas (TDG) supersaturation downstream of dams can occur in the Yangtze River basin and is known to cause stress and even death in fish. Consequently, it is important to establish tolerance thresholds of endemic fish to protect local aquatic resources. We conducted experiments to assess survival characteristics and swimming ability of bighead carp, an important commercial fish dwelling in the Yangtze River, to evaluate its tolerance threshold to TDG supersaturation. The typical external symptoms of gas bubble trauma (GBT) were observed and the time when the fish lost equilibrium and died were recorded. The results showed that the mortality occurred when TDG level exceeded 125%, with obvious symptoms such as exophthalmos and bubbles on the head. The interval between loss of equilibrium and mortality decreased with an increase in TDG level. Neither exposure time nor TDG level significantly affected the critical swimming speed (Ucrit) of fish exposed to non-lethal exposure (110%, 120% and 125% TDG) over a 7 day period. Significant reductions in Ucrit were found under 130% and 135% TDG conditions when the exposure lasted 52.0 h and 42.9 h, respectively. The Ucrit also significantly decreased after exposure of 1.6 h under 140% TDG condition. Moreover, after exposure to 140% TDG for 39.2 h, 135% TDG for 56.5 h and 130% TDG for 95.9 h, bighead carp were transferred into air saturated water to recover for 24 h or 48 h; however, swimming performance remained impaired. The results of this study indicate that 125% TDG was the highest TDG level where limited mortality was observed and the swimming ability was not impaired, showing that 125% TDG can be set as the tolerance threshold of this species to guide the operation of dams in the Yangtze River Basin.

Fractionation of metals in soil for strawberry cultivation: Effect on metal migration in food chain and application in risk assessment.

Although trace metals in strawberry production system have attracted growing attention, little is known about metal fractionation in soil for strawberry cultivation. We hypothesized that the metal fractions in soil influenced by strawberry production had significant effect on food chain transport of metals and their risk in soil. Here, samples of strawberries and soil were gathered in the Yangtze River Delta, China to verify the hypothesis. Results showed that the acid-soluble Cr, Cd, and Ni in soil for strawberry cultivation were 21.5%-88.3% higher than those in open field soil, which enhanced uptake and bioaccessible levels of these metals in strawberries. Overall, the ecological, mobility, and health risks of Pb, Zn, Ni, and Cu in soil were at a low level. However, the ecological risk of bioavailable Cd, mobility risk of Cd, and cancer risk of bioavailable Cr in over 70% of the soil samples were at moderate, high, and acceptable levels, respectively. Since the increased acid-soluble Cr and Ni in soil were related to soil acidification induced by strawberry production, nitrogen fertilizer application should be optimized to prevent soil acidification and reduce transfer of Cr and Ni. Additionally, as Cd and organic matter accumulated in soil, the acid-soluble Cd and the ecological and mobility risks of Cd in soil were enhanced. To decrease transfer and risk of Cd in soil, organic fertilizer application should be optimized to mitigate Cd accumulation, alter organic matter composition, and subsequently promote the transformation of bioavailable Cd into residual Cd in soil.

Effect of total dissolved gas supersaturation and flow velocity on survival and swimming ability of juvenile Schizothorax prenanti.

Although developing large-scale hydropower cascades in the upper Yangtze River effectively improves the hydropower resource utilization, it produces total dissolved gas (TDG) supersaturation. In the flood season, the high level of TDG supersaturation (TDGS) frequently occurs in the downstream of dams, causing migratory fish to suffer from gas bubble trauma (GBT) and reducing their survival and swimming ability. Currently, there is a deficiency in particular approaches to evaluate the ecological hazard posed by TDGS on migratory fish as they traverse different flow velocities within their migratory routes. This study assessed the vulnerability of juvenile Schizothorax prenanti (S. prenanti) to GBT from the static setting to 9.0 BL/s during exposure to nominal levels of 100%, 110%, 120% and 130% TDG. The mortality occurs when the flow velocity surpasses 6.0 and 7.5 BL/s in 100% and 110% TDG levels, respectively. For fish exposed to 120% and 130% TDG levels, the relationship between survival time and flow velocity is an approximately inverse bell-shaped curve with increasing velocity. The optimal velocity of maximal survival time of juvenile S. prenanti is 3.0 and 4.5 BL/s in 120% and 130% TDG-supersaturated water. Both TDG level and flow velocity significantly affect burst swimming speed (Uburst) and critical swimming speed (Ucrit). The cases involving GBT showed substantial declines in Uburst and Ucrit, exceeding 6.0 BL/s and TDG levels greater than 120%. The results may contribute to formulating a specific management strategy for hydropower operation during the migratory period and conserving vulnerable species in the Yangtze River.

Mortality risk evaluation methods for total dissolved gas supersaturation to fish based on a mitigation measure of utilizing activated carbon.

The proper water chemical composition of aquaculture water is very important for fish farming in reservoirs or fish multiplication stations. Gas bubble disease (GBD) is mainly caused by total dissolved gas supersaturation (TDGS) in water and is a common problem that affects the healthy growth of fish. Extensive measures have been taken to mitigate TDGS levels in water where fish live, while methods for quantitatively evaluating the mitigation effect of the proposed measures on fish exposed to TDGS are still lacking. In this paper, an activated carbon (AC) adsorption experiment for supersaturated total dissolved gas (TDG) dissipation was conducted, and the experimental results indicated that AC addition could effectively accelerate supersaturated TDG dissipation. Based on fish tolerance experiments conducted by Huang (2010), two models, including a mortality risk degree evaluation model and a mortality rate calculation model, were developed to quantitatively evaluate the mortality risk mitigation effect of AC addiction on fish exposed to unsteady TDGS levels. Application of the results of the mortality risk degree evaluation model has shown that AC addition can help alleviate the mortality risk of fish suffering from TDGS. Application of the results of the mortality rate calculation model has also demonstrated that the final mortality rate of the fish group in the case with AC addition was lower than that of the case without added AC, and the final mortality rate decreased as the specific surface area and dosage of AC increased. Furthermore, an equation that related the required AC mass and a given harvested fish mass was established. This paper provides a reference for evaluating the effects of various mitigation measures to alleviate the risk posed to fish by TDGS.

Production of total dissolved gas supersaturation at hydropower facilities and its transport: A review.

Total dissolved gas supersaturation (TDG) is a common issue in hydropower facilities as a result of water conveyance structures that increase the amount of air entrainment from the atmosphere and dissolved into the water. Water with TDG supersaturation can negatively impact fish, aquatic invertebrates and their habitats. This study comprehensively reviewed the physical mechanisms of TDG generation and predictive TDG generation models at various facility types. To establish TDG mitigation strategies, it is essential to develop predictive tools for TDG generation that consider both facility geometry as well as the hydrology of the downstream environment. Applications of TDG prediction at different discharge modes included plunging flows, trajectory jets, plunging jets, free-falling jets, and submerged jets were discussed. TDG transport models in downstream rivers involving mixing and dissipation were introduced, which can be integrated with TDG generation models into a platform to describe TDG distribution in river systems. Subsequently, risk ranking procedures for assessing the degree of TDG risk on fish were provided. Potential measures for mitigating TDG supersaturation were reviewed and included engineering, operational, and technical solutions. Outcomes from this review considered a diverse suite of studies on TDG issues in regulated rivers and allowed for recommendations to reduce uncertainties and improve environmental performance at facilities where TDG risks occur.

Swimming performance of a pelagic species in the Yangtze River under different exposure modes of the total dissolved gas supersaturation.

During flood discharges of upstream dams in the Yangtze River, the pelagic fish have a stress risk from total dissolved gas (TDG) supersaturation in the river water. This study took the silver carp as the object and systematically evaluated the effects of TDG supersaturation levels and exposure time on their critical swimming speed (Ucrit) at different temperatures. The external symptoms of gas bubble disease were found when TDG levels exceeded 130%. Both exposure time and TDG level did not significantly impact the Ucrit of fish under 6 days of non-lethal exposure (110%, 120%, 130% TDG) with lower or higher water temperature. Significant differences in Ucrit were found among different exposure times at 11.0 ± 1.0°C under 10 hours of lethal exposure (135%, 140%, 150% TDG) and the Ucrit reduced by 59.88%, 83.32%, and 92.40%, respectively. TDG level had a significant impact on the Ucrit at 21.0 ± 1.0°C when exposure time exceeded 8 hours. Ucrit at 21.0 ± 1.0°C water were significantly greater than those at 11.0 ± 1.0°C water where conditions had the same TDG supersaturation and exposure time. Differences in Ucrit between temperatures ranged from 3.24 to 6.12 BL/s under non-lethal exposure and from 6.38 to 13.88 BL/s under lethal exposure. The results of this study can provide a reference for fish conservation during flood discharge.

River habitat assessment and restoration in high dam flood discharge systems with total dissolved gas supersaturation.

The success of river habitat restoration relies on accurate assessment proxies. However, determining how to quantitatively assess the impact of multiple stressors during flood discharge from high dams in riverine ecosystems and where and how to implement more reliable recovery interventions remain challenges. Here, we developed a bottom-up mechanistic framework for assessing the effects of total dissolved gas supersaturation (TDGS) and hydrodynamics on fish habitat quality and applied it to the downstream river reach of the Xiangjiaba Dam in Southwest China. The results showed that the available habitat area of river sturgeon was the smallest, while Chinese sucker had the largest available habitat area among the three target species under all discharge scenarios. Although the TDGS levels were evenly mixed laterally, the habitat suitability index indicated that the suitable habitats were primarily within both sides of the river reach under all scenarios, which is contrary to findings based on the traditional TDGS risk assessment model. The traditional TDGS risk assessment model overestimates the impact of dams on habitats. This divergence reflected the sensitivity of the habitat assessment to fish habitat preferences, fish tolerance to TDGS and the biological response of fish under TDGS. Additionally, the priority areas for restoration can be identified by habitat suitability index with lower values. We simulated twenty-four schemes and found that interventions such as stone groups, ecological spur dike, water-retaining weir and river dredging can enhance habitat suitability for fish species under multiple stressors, providing novel insights into where and how to mitigate the impact of TDGS. Our findings offer a transferable framework for the quantitative evaluation of fish habitat and implementation of restoration management during dam flood discharge periods, thus providing a new perspective for biodiversity conservation and habitat restoration in dam-regulated rivers with TDGS around the world.

Direct identification of A-to-I editing sites with nanopore native RNA sequencing.

Inosine is a prevalent RNA modification in animals and is formed when an adenosine is deaminated by the ADAR family of enzymes. Traditionally, inosines are identified indirectly as variants from Illumina RNA-sequencing data because they are interpreted as guanosines by cellular machineries. However, this indirect method performs poorly in protein-coding regions where exons are typically short, in non-model organisms with sparsely annotated single-nucleotide polymorphisms, or in disease contexts where unknown DNA mutations are pervasive. Here, we show that Oxford Nanopore direct RNA sequencing can be used to identify inosine-containing sites in native transcriptomes with high accuracy. We trained convolutional neural network models to distinguish inosine from adenosine and guanosine, and to estimate the modification rate at each editing site. Furthermore, we demonstrated their utility on the transcriptomes of human, mouse and Xenopus. Our approach expands the toolkit for studying adenosine-to-inosine editing and can be further extended to investigate other RNA modifications.

Constructing nickel cobaltate @nickel-manganese layered double hydroxide hybrid composite on carbon cloth for high-performance flexible supercapacitors.

Flexible supercapacitors have received considerable interest owing to their potential application in wearable electronics. Designing subtle hybridization of active materials and constructing smart electrode architectures are effective strategies for developing high-performance flexible supercapacitors. Herein, a hierarchically hybrid electrode is engineered by integrating nanoneedle-like structural NiCo2O4 and NiMn layered double hydroxide (NiMn-LDH) composite on highly conductive carbon cloth (CC). This architecture can endow abundant active sites, rapid electron collection pathways and efficient ion transport channels. The resultant hybrid electrode delivers high areal capacitance of 4010.4 mF cm-2, excellent cyclic stability and good rate performance. Furthermore, by pairing with an activated carbon (AC)/CC anode, a flexible solid-state asymmetric supercapacitor (ASC) is assembled, which exhibits the high areal energy/power density of 0.78 mWh cm-2/40.4 mW cm-2 and superior capacitive stability at bending deformation. Meanwhile, the assembled ASC possesses outstanding cycling stability with 97.7% capacitance retention after 10,000 cycles. This work presents the effects of rational design of hybrid electrode with high electrochemical properties and flexibility, holding great potential for flexible energy storages.

Effects of Total Dissolved Gas Supersaturation on the Survival of Juvenile Procypris rabaudi and Juvenile Myxocyprinus asiaticus at Varying Water Depth in a Natural River.

Total dissolved gas (TDG) supersaturation, which can be caused by flood discharge, results in gas bubble disease (GBD) in fish and threatens their survival downstream of dams. TDG supersaturation has become a serious environmental problem in the Yangtze River. Few studies have evaluated the effect of TDG supersaturation on fish in natural rivers during periods of flood discharge. To estimate fish tolerance to TDG supersaturation under natural conditions, juvenile Myxocyprinus asiaticus and juvenile Procypris rabaudi were exposed to TDG-supersaturated water for 96 h at various depths (0-0.3 m, 0.3-1.3 m, 1.3-2.3 m and 0-2.3 m) during periods of flood discharge of Dagangshan hydropower station. The results showed that juvenile Procypris rabaudi and juvenile Myxocyprinus asiaticus exhibited obvious GBD signs. An increase in exposure time decreased survival probability of the two species. Deeper water depths can increase the tolerance of juvenile Procypris rabaudi to TDG supersaturation in natural rivers during periods of flood discharge while it cannot improve the survival of juvenile Myxocyprinus asiaticus. Compared with juvenile Myxocyprinus asiaticus, juvenile Procypris rabaudi showed weaker tolerance of TDG supersaturation in shallow water, and juvenile Procypris rabaudi were more vulnerable to TDG supersaturation than juvenile Myxocyprinus asiaticus even if the TDG level (116%) was low.

Microplastics in inland freshwater environments with different regional functions: A case study on the Chengdu Plain.

As an emerging pollutant, microplastic has been demonstrated to widely cover water and sediments in marine and freshwater environments globally. Due to the differences in the regional functions of cities, the abundance and characteristics of microplastic pollution in rivers are different. Taking the Minjiang River as the research object, which flows through the Chengdu Plain, the abundance, composition, shape, and size of microplastics in the water and sediments of the Minjiang River were investigated. The results showed that serious microplastic pollution occurred in the Chengdu section of the Minjiang River, with an abundance ranging from 6.11-44.08 n/L in the water and from 573.84-2878.97 n/kg in the sediments. By using the "regional function index (Q)", the relationships between the characteristics of microplastic pollution and regional functions were analyzed. Areas with a high Q proved to be more polluted by microplastics. Densely populated areas with large construction areas and high human activity intensity tended to generate microplastics with larger sizes and a more fiber-like shapes. Rayon (RA) and nylon (PA) were identified as typical urban microplastics, while polypropylene (PP) and polyethylene (PE) were identified as typical agricultural microplastics. This study elucidated the microplastic pollution in the Chengdu section of the Minjiang River, a tributary in the upper reaches of the Yangtze River. It also provided a new direction for the study of microplastic pollution characteristics of freshwater environments with different regional functions and microplastic pollution source control.

Interleukin-33 Promotes Th2/Th17 Response in Eosinophilic and Non-Eosinophilic Nasal Polyps.

BACKGROUND: Interleukin-33 (IL-33) is reported to be involved in Th2-skewed eosinophilic inflammation. A recent study also found that IL-33 exerted opposite effects on Th17 response in different diseases. However, the role of IL-33 in chronic rhinosinusitis with nasal polyps (NPs) was not explored. OBJECTIVES: The purpose of this study was to investigate the expression and function of IL-33 in chronic rhinosinusitis with NPs. MATERIALS AND METHODS: NP tissues from 60 NP patients and normal tissues of the inferior turbinate from 20 controls were sampled in operation. Immunochemistry was performed to identify eosinophilic or non-eosinophilic NPs. The expressions of IL-33 and Th1/2/17 cytokines were compared between different subtypes of NPs. The effect of IL-33 on Th response was detected in dispersed nasal polyp cells (DNPCs), and the signaling pathways involved in the process were detected using Western blot. RESULTS: The concentration of IL-33 was significantly elevated in both eosinophilic and non-eosinophilic NPs compared with controls. By in vitro study, we found that IL-33 can induce IL-4 and IL-5 production from eosinophilic DNPCs through the PI3K/AKT pathway, whereas IL-33 can induce IL-17 production from non-eosinophilic DNPCs through the ERK1/2 pathway. CONCLUSION: IL-33 is involved in Th2/Th17 response in NPs. Our study suggests that different types of NPs need a different treatment target.

Effects of Total Dissolved Gas Supersaturation in Fish of Different Sizes and Species.

Two endemic fish in the upper Yangtze River, the Rock Carp (Procypris rabaudi) and Prenant's Schizothoracin (Schizothorax prenanti), were used as research objects in this study to assess the effects of total dissolved gas (TDG) supersaturation on fish of varying sizes. Fish were exposed to TDG-supersaturated water at the levels of 145, 140, 135, 130, and 125%. The results showed that fish swam slowly, responded clumsily, and then exhibited spiral swimming performance after a period of exposure to TDG-supersaturated water. Fish exhibited exophthalmos, body swelling, gill bleeding, and caudal fin bleeding when they died in the TDG-supersaturated water. With the increase in TDG supersaturation, the tolerance capacity of fish to supersaturated TDG significantly reduced. At high supersaturation, the difference in survival time between species was not significant, while fish with smaller sizes showed greater tolerance capacity. At low supersaturation, the tolerance capacity of fish was mainly affected by species, and the influence of size was relatively small. With the decrease in TDG supersaturation, the catalase (CAT) activity first increased and then decreased. Rock Carp displayed significantly less activity than Prenant's Schizothoracin on exposure to TDG-supersaturated water. At high supersaturation levels, the CAT activity of Prenant's Schizothoracin of small size was greater than that of large Prenant's Schizothoracin. In contrast, small Prenant's Schizothoracin showed less CAT activity at low TDG levels than did large individuals.

The Tolerance Characteristics of Resident Fish in the Upper Yangtze River under Varying Gas Supersaturation.

In circumstances where total dissolved gas (TDG) levels are variable, the peak TDG and duration are expected to be the dominant drivers of fish survival. Focusing on the peak TDG and duration in natural rivers, a laboratory experiment and field experiments in the upper Yangtze River were conducted with Prenant's Schizothoracin (Schizothorax prenanti), a rare species inhabiting the upper Yangtze River, to examine the tolerance characteristics of fish under varying gas supersaturation levels. The results of the field experiments showed that TDG supersaturation in natural rivers changed greatly during the flood period due to reservoir regulation. The survival of fish was affected by TDG levels, water depth and TDG fluctuation range. A high TDG level, and shallow compensatory water depth caused fish mortality in the field experiment to be higher in September than in July. The results of the laboratory experiment showed that fish tolerance was lower under fluctuating TDG supersaturation than under constant TDG supersaturation. The tolerance of fish to TDG supersaturation varied depending on peak TDG and duration. Under the fluctuation range of 115-125%, fish survival in the 6 h-6 h cycle was significantly different from that in the 8 h-8 h cycle. The fluctuation cycle did not affect fish survival at the fluctuation range of 110-130%. Intermittent lower TDG supersaturation does not significantly increase the tolerance of fish. This study revealed the tolerance characteristics of resident fish in the upper Yangtze River to TDG supersaturation, which provides a reference to the ecological operation of reservoirs and may contribute to the protection of aquatic organisms.

Effects of water depth on GBD associated with total dissolved gas supersaturation in Chinese sucker (Myxocyprinus asiaticus) in upper Yangtze River.

Spillway water falling from hydroelectric power plant dams in the upper Yangtze River creates a high pressure in plunge pools below the dams allowing gasses to be dissolved at high rates. The resulting supersaturation persists many miles downstream the dam which may elicit mortality in river fishes associated with gas bubble disease (GBD). We have in a two-year study (2014-15) evaluated the effect of water depth on development of GBD in an endemic and endangered fish species, the Chinese sucker Myxocyprinus asiaticus, 24 km downstream of Xiangjaiba dam. Mortality and incidence of GBD were recorded and it was seen that water depth and survival time/GBD development was positively correlated. The physiological mechanisms explaining increased resistance to GBD with increased water depths (and thereby higher hydrostatic pressure) are discussed. The results may be applied in future management of fish resources in order to protect endangered endemic fishes in rivers affected by dam constructions.

Tailoring Three-Dimensional Composite Architecture for Advanced Zinc-Ion Batteries.

Rechargeable aqueous Zn-ion batteries (ZIBs) are of considerable interest for future energy storage. Their main limitation, however, is developing suitable cathode materials capable of sustaining the Zn2+ repeated intercalation/deintercalation. Herein, a three-dimensional polypyrrole (PPy)-encapsulated Mn2O3 composite architecture is developed for advanced ZIBs. The engineering can be easily realized via in situ phase transformation of MnCO3 microboxes with subsequent self-initiated polymerization of PPy. The abundant open-up pores (∼30 nm) throughout the construction accelerate ion migration and provide a more active interface for Zn2+ storage in the Mn2O3@PPy bulk electrode. Meanwhile, the PPy skin uniformly wrapped on the Mn2O3 microbox not only guarantees a good conductive network for faster electron transport but also inhibits the dissolution of Mn2O3 and protects the integrity of the electrode from structural damage. As a result, the Mn2O3@PPy electrode can operate at reversible capacity exceeding those of most other cathode materials, but can still provide longer lifetime (no capacity decay over 2000 cycles at 0.4 A g-1) and higher rate performance than others. Furthermore, theoretical studies show the H+ and Zn2+ coinsertion storage mechanism and reaction dynamics. The results show that this three-dimensional Mn2O3@PPy architecture is a promising cathode material for high-performance ZIBs.

Effects of Total Dissolved Gas Supersaturation on the Swimming Performance of Two Endemic Fish Species in the Upper Yangtze River.

Total dissolved gas (TDG) supersaturation has been identified as one of the possible negative environmental effects of the construction of dams in the upper Yangtze River. Juvenile Chinese sucker and Prenant's schizothoracin fish were selected to evaluate the impact of TDG supersaturation on the swimming performance of fish in the Upper Yangtze River. The critical swimming speeds (Ucrit) of Chinese sucker were 4.06, 2.83, 2.87, 2.68, and 2.29 BLs-1 at the TDG supersaturation levels of 100, 117, 122, 125 and 130%, respectively. The Ucrit of Prenant's schizothoracin were 7.38, 4.32, 3.98, and 3.74 BLs-1 at the TDG supersaturation levels of 100, 117, 125 and 130%, respectively. The burst swimming speed (Uburst) of the two species also significantly declined with increases in the TDG supersaturation level. The present results demonstrate that the swimming speeds of Prenant's schizothoracin that were exposed to 130% TDG supersaturation for 2 h exhibited significant recovery after 2 days, whereas the swimming speeds of Chinese sucker did not. The swimming speeds of Chinese sucker after 2 days of recovery were significantly reduced compared with those of control fish, whereas the speeds of Prenant's schizothoracin returned to normal levels.

[SiRNA silencing Rsk2 gene expression inhibits proliferation and osteogenic differentiation of human periodontal ligament cells].

PURPOSE: To study the effect of ribosomal S6 kinase (Rsk2) gene on proliferation and osteogenic differentiation of human periodontal ligament cells (hPDLCs) and underlying mechanism. METHODS: Premolar surgically extracted were collected, the periodontal ligament was separated and hPDLCs were primarily cultured. Cells of 4 to 6 passage were used in the experiment. The silencing efficiency of small interfering RNA (siRNA) on Rsk2 in hPDLCs was detected by RT-PCR and Western blot. MTT assay was used to detect the effect of Rsk2 siRNA on cell proliferation. Alkaline phosphatase (ALP) kit was used to detect ALP activity. P38MAPK signal pathway inhibitor SB203580 was used to detect hPDLCs after transfection. Western blot was used to detect the effect of Rsk2 siRNA on MAPK signaling pathway p38 protein phosphorylation. The expressions of Runt-related transcription factor-2 (Runx2), osteocalcin (OCN) and osteogenic protein BMP2 were detected by RT-PCR and Western blot. The data were analyzed using SPSS18.0 software package. RESULTS: The expression of Rsk2 was down-regulated by hPDLCs transfected with Rsk2 siRNA, the difference was statistically significant (P<0.05). Rsk2 siRNA significantly reduced phosphorylation of p38 protein (P<0.05), inhibition of hPDLCs proliferation (P<0.05), decreased ALP activity (P<0.01); the expression of Runx2, OCN and BMP2 was different, and the difference was statistically significant (P<0.05). After SB203580 treatment, hPDLCs transfected with Rsk2 siRNA showed increased cell proliferation, ALP activity, Runx2, OCN and BMP2 expression; compared with Rsk2 siRNA transfected hPDLCs, the difference was statistically significant. CONCLUSIONS: Rsk2 siRNA inhibits hPDLCs proliferation and osteogenic differentiation through p38MAPK signaling pathway.

A chemical-inducible CRISPR-Cas9 system for rapid control of genome editing.

CRISPR-Cas9 has emerged as a powerful technology that enables ready modification of the mammalian genome. The ability to modulate Cas9 activity can reduce off-target cleavage and facilitate precise genome engineering. Here we report the development of a Cas9 variant whose activity can be switched on and off in human cells with 4-hydroxytamoxifen (4-HT) by fusing the Cas9 enzyme with the hormone-binding domain of the estrogen receptor (ERT2). The final optimized variant, termed iCas, showed low endonuclease activity without 4-HT but high editing efficiency at multiple loci with the chemical. We also tuned the duration and concentration of 4-HT treatment to reduce off-target genome modification. Additionally, we benchmarked iCas against other chemical-inducible methods and found that it had the fastest on rate and that its activity could be toggled on and off repeatedly. Collectively, these results highlight the utility of iCas for rapid and reversible control of genome-editing function.