Our results show that a small amount of Rb+ was enough to induce the crystallization of the α-FAPbI3 phase and suppress the formation of the yellow non-photoactive phase; the grain size increased, and the product of the carrier mobility and the lifetime (μτ) improved. As a result, the fabricated photodetector exhibited a broad photo-response region, from ultraviolet to near-infrared, with maximum responsivity (R) up to 11.8 mA W−1 and excellent detectivity (D*) values up to 5.33 × 1011 Jones. This work provides a feasible strategy to improve photodetectors’ performance via additive engineering.Photodetectors are widely employed as fundamental devices in optical communication, automatic control, image sensors, night vision, missile guidance, and many other industrial or military fields. Mixed-cation perovskites have emerged as promising optoelectronic materials for application in photodetectors due to their superior compositional flexibility and photovoltaic performance. However, their application involves obstacles such as phase segregation and poor-quality crystallization, which introduce defects in perovskite films and adversely affect devices’ optoelectronic performance. The application prospects of mixed-cation perovskite technology are significantly constrained by these challenges. Therefore, it is necessary to investigate strategies that combine crystallinity control and defect passivation to obtain high-quality thin films. In this study, we incorporated different Rb The family of all-inorganic perovskites has fewer members owing to the limitations of the tolerance factor and octahedral factor [6]4−. Numerous chemically versatile multidimensional OIHP structures can be obtained by modifying the A-site organic cation component due to the wide variety of organic cation groups. OIHPs can adapt to various optoelectronic applications such as solar cells, photodetectors, and light-emitting diodes [3) has been extensively investigated and exhibits outstanding photoelectric conversion efficiency as a potential absorber in solar cells [3 crystal structure [3 has a tetragonal rather than cubic structure at room temperature, with a band gap of 1.51 eV due to the small size of the MA ion [3, formamidinium lead iodide (FAPbI3) possesses higher symmetry and is closer to a cubic structure [3 exhibits superior thermal stability and a favorable band gap closer to the optimal band (1.4 eV) [3 one of the most promising absorbing layers for high-performance photodetectors [3 perovskite has only two phases at room temperature: one is a photoactive cubic phase (α-phase), and the other is a non-photoactive hexagonal phase (δ-phase) [3 without the δ-phase and stabilizing the α-phase have become urgent problems. In particular, mixed-cation engineering has been demonstrated as an effective strategy for improving perovskites’ phase stability. Jeon et al. proved that a small amount of MA+ is enough to induce improved crystallization of black α-FAPbI3 [+ can effectively suppress the formation of yellow-phase impurities in MA/FA perovskite compounds; a device prepared based on a Cs/MA/FA triple-cation yielded stabilized power-conversion efficiency exceeding 21% [− and Br−) to form different halide compositions, which incorporate various impurities and defects into the film [+ or Rb+ in triple-cation perovskites has been proven to alleviate lattice strain, which is beneficial for suppressing the formation of FA+ and I− vacancies [+ does not dope the lattice of Cs/MA/FA perovskite, avoiding the introduction of extra lattice strains. Instead, Rb+ may form various Rb-containing compounds with excess PbI2 and can act as a potential passivation layer, which is beneficial for decreasing non-radiative recombination and improving carrier transport [In the past decade, solution-processed organic–inorganic hybrid perovskites (OIHPs), which can be prepared by a simple and low-cost spin coating method, have garnered widespread attention in the field of optoelectronics due to their superior properties, such as high absorption coefficient, high carrier mobility, low exciton binding energy, and adjustable band gap ,2,3. In l factor . Compareg diodes ,6,7. OIHg diodes ,9. In paar cells ,11,12. Htructure . Moreovee MA ion . Comparetructure ,16. Furt(1.4 eV) . The redetectors ,19,20. Gδ-phase) . Unfortuα-FAPbI3 . Furtherding 21% . Neverthding 21% ,24,25. Pthe film . It is wthe film . To addrthe film . For exathe film . F4TCNQ the film ,31. Receacancies ,33. Moreransport . These f+ into the triple-cation perovskite precursor solution Cs0.05(MA0.17FA0.83)0.95Pb(I0.83Br0.17)3 (Cs0.05(MA0.17FA0.83)0.95 is abbreviated as CsMAFA in this paper). We demonstrated via X-ray diffraction and scanning electron microscopy that the optimal ratio of Rb+ can improve crystallinity and suppress the formation of non-photoactive phases. Incorporating Rb+ increased the grain size and reduced grain boundaries, which contributed to the inhibition of non-radiative complexation and improved carrier mobility. Therefore, the fabricated photodetector exhibited a broad photo-response region from ultraviolet (UV) to near-infrared (NIR) with a maximum responsivity of up to 11.8 mA W−1 and an excellent D* value of up to 5.33 × 1011 Jones. The maximum switching ratio was 7252, and the rise and decay times were 208.3 μs and 288.4 μs, respectively. During measurement under ambient light, no significant photocurrent decays were found during multiple cycles.In this study, we incorporated different ratios of Rb2, 99.0%, Acros, Geel, Belgium) were purchased from Beijing Innochem Science & Technology Co., Ltd., Beijing, China. Lead (II) iodide  was purchased from Xi’an Polymer Light Technology Corp., Xi’an, China. Super-dehydrated dimethylformamide  was purchased from Beijing J&K Scientific Technology Corp., Beijing, China. Dimethyl sulfoxide  was purchased from Alfa Aesar Chemical Corp., Taiwan, China. All chemicals were purchased from commercial suppliers and used without further processing.Formamidinium iodide , cesium iodide , methylammonium bromide , and lead (II) bromide (PbBr2 layer was deposited from a precursor of titanium diisopropoxide (0.6 mL) and bis(acetylacetonate) (0.4 mL) in anhydrous propan-2-ol (7 mL). The precursor solution was sprayed onto the glass substrate, which was subsequently annealed on a hot plate at 460 °C for an hour and then left to cool to room temperature. The Cs/MA/FA perovskite precursor solution was prepared by dissolving 1 M FAI, 1.1 M PbI2, 0.2 M MABr, 0.063 M CsI, and 0.22 M PbBr2 in anhydrous DMF/DMSO 4:1 (v/v). Next, the quadruple cation was prepared by adding 0%, 5%, and 10% of the 1.0 M RbI solution to the Cs/MA/FA perovskite precursor solution. The quadruple-cation precursor solution was spin-coated onto the glass/c-TiO2 substrate at 1500 rpm for 5 s and then at 4000 rpm for 30 s; the glass/c-TiO2/perovskite substrate was subsequently annealed on a hot plate at 100 °C for 60 min. A 60 nm gold interdigitated electrode (distance between electrodes: L = 50 µm) was deposited via thermal evaporation under a pressure of 4 × 10−4 Pa. A black mask was pasted on to guarantee a standard illumination area of 5 mm2.The glass substrates were cleaned twice with deionized water, and then with ethanol. After each cleaning, the glass substrates were sonicated for 20 min. After sonication, the glass substrates were blown dry with nitrogen, and then a UV–ozone treatment was performed for 30 min. A compact TiO−1). The morphologies of thin films were measured using a SU8010 SEM . The photodetector’s performance was measured using an LED light source with different wavelengths. All current data were recorded using an electrochemical workstation with a CIMPS system .UV–vis spectra were measured using the UV–2450  from 400 nm to 900 nm. XRD patterns were obtained with an X-ray diffractometer  using Cu–Kα radiation (1.5418)  perovskites have achieved impressive high-efficiency solar cells (25.7%) and become a popular material in the optoelectronic devices field. However, FAPbI3’s octahedral skeleton is unstable owing to the lattice distortion caused by FA ions, which may form vacancies in the perovskite lattice. According to the principle of mechanical force, internal stresses occur and resist this effect when large cations stretch the octahedral skeleton, leading to an unstable state in the whole system [+ and Cs+, at the A-site [+, MA+, and Br− into the FA-based perovskite’s lattice, yielding a higher symmetry.Possessing a broad optical absorption spectrum and high absorption coefficient, black-phase FAPbIe system . Figure e system . One stre A-site . Figure + incorporation on the growth of CsMAFA perovskite films, X-ray diffraction measurements were taken. As shown in +) film, a typical characteristic peak appeared at 14.06°. The main reflection peak of α-FAPbI3 (14°) shifted to a higher angle, indicating that the doping of Cs+ caused the contraction of the MAFA-based perovskite lattice, which facilitated the release of lattice strain. Moreover, we noted small side peaks at 11.56° and 12.96°, corresponding to non-photoactive (δ-FAPbI3) and excess PbI2, respectively. This was a typical observation of the incomplete conversion of mixed-cation perovskites to the photoactive phase. Subsequently, when we introduced 5% of Rb+, the main reflection peak further shifted to 14.10° and the peak intensity increased, demonstrating that the introduction of Rb+ could further alleviate lattice distortion in the CsMAFA-based perovskite. In particular, the intensity of the δ-phase diminished when we introduced Rb+, and 5% of Rb+ suppressed the formation of the δ-phase most effectively, which indicated that a small amount of Rb+ was able to promote the CsMAFA-based perovskite crystallizing to the α-phase. Notably, the decrease in the small side peak at 12.96° may suggest that Rb+ consumed the residual PbI2 reaction to form the Rb-containing mixture [+ may compete with the incorporation of Cs+ into the perovskite lattice by forming a stable mixed Cs/Rb hexagonal phase.To further investigate the effect of Rb mixture . Moreove+ concentration was increased to 10%. Therefore, excessive Rb+ may be detrimental to the crystallization of CsMAFA-based perovskite. In terms of crystal quality, the optimal ration of Rb+ was 5%.The main reflection peak shifted slightly to 14.08° and the diffraction peak’s intensity decreased when the Rb0.05(CsMAFA)0.95 were investigated using scanning electron microscopy (SEM) (0.05(CsMAFA)0.95 film improved from 211 nm to 226 nm, which indicates that Rb+ promoted crystallization and matched the XRD result. Energy Dispersive Spectroscopy (EDS) on the surface of the Rb 0.05(CsMAFA)0.95 film also confirmed the presence of Rb+, as shown in + to the perovskite precursor solution could effectively control the crystallization process of the film, which was beneficial for yielding highly crystalline and crack-free perovskite films to improve photodetectors’ performance.It is well known that films with a large grain size and fewer grain boundaries are essential for reducing carrier recombination and enhancing carrier mobility, as pinholes or cracks tend to form non-radiative recombination centers that adversely affect devices’ performance. The surface morphologies of CsMAFA and Rbpy (SEM) a,b. As s+ ratios. The CsMAFA film has a strong absorbance in the region of wavelengths less than 450 nm, and the absorption band could extend beyond 800 nm. The Rb0.05(CsMAFA)0.95 film’s absorbance significantly improved in the region of wavelengths less than 752 nm, with a band gap of 1.65 eV calculated using the Tauc formula  product. To quantitatively characterize the effect of different Rb+ ratios on the detector’s charge collection efficiency, we fabricated three devices separately and performed photoconductivity measurements. In this paper, we used a gold interdigitated electrode to collect carriers, as shown in µτ values. The Hecht equation is expressed as follows [I0 is the saturated photocurrent and d is the distance between electrodes [µτ values of CsMAFA, Rb0.05(CsMAFA)0.95, and Rb0.1(CsMAFA)0.9 were 8.37 × 10−6 cm2 V−1, 1.03 × 10−5 cm2 V−1, and 9.01 × 10−6 cm2 V−1, respectively. The Rb0.05(CsMAFA)0.95 device yielded the highest µτ value, further demonstrating the beneficial effect of small amounts of Rb+ on the crystallization of films. −2 and a bias of 4 V. The calculated on–off values for CsMAFA, Rb0.05(CsMAFA)0.95, and Rb0.1(CsMAFA)0.9 were 64, 1519, and 37, respectively. Notably, the photocurrent of the Rb0.1(CsMAFA)0.9 device decreased instead, which could probably be attributed to cesium’s tendency to form a stable Cs0.5Rb0.5PbI3 phase in the presence of excess rubidium [+ could yield the highest switching ratio. Based on the above analysis, we determined that the optimal Rb+ ratio was 5% and measured the current–voltage (I–V) characteristics (R) is an important parameter characterizing the ability of a photodetector to convert photons into a current signal, and it can be calculated using the following equation [pI is the photocurrent, dI is the dark current, P is the light intensity, and S is the effective area of the device. R versus light intensity. The photocurrent increased linearly with increasing light intensity under 454 nm illumination, whereas the responsivity decreased with increasing light intensity due to the dominance of bimolecular recombination under strong light [R was 11.8 mA W−1 at a light intensity of 0.5 mW cm−2. Linear dynamic range (LDR) is a figure-of-merit that represents the range of light intensity over which a detector can output photocurrent linearly. LDR can be calculated using the following formula [maxP is the saturation light intensity under which the photocurrent begins to deviate from linearity and minP is the initial light intensity. −2 to above 20 mW cm−2, yielding an LDR of 32 dB. We further evaluated the specific detectivity, which is a parameter characterizing a detector’s ability to detect weak light and is calculated using the following expression:R is the responsivity, e is the unit charge, dI is the dark current, and S is the active area. D* with light intensity. The maximum value of D* was 5.33 × 1011 Jones under light irradiation at 454 nm, 0.5 mW cm−2, which proves its potential application in weak near-ultraviolet light sensing and imaging. External quantum efficiency (EQE) is also a key photodetector parameter and is defined as the ratio of the number of electron–hole pairs collected by the external circuit to the number of incident photons. It is calculated as follows:R is the responsivity, h is Planck’s constant, c is the speed of light, e is the elementary charge, and λ is the wavelength of the incident light. The EQE versus wavelength is shown in EQE reached a peak of 1.5% at a wavelength of 454 nm, with a D* up to 2.49 × 1011 Jones due to the high absorption coefficient in the near-UV band. The EQE gradually decreased in the region of wavelengths above 632 nm; the lowest D* was 9.38 × 108 Jones at 868 nm.Detectors’ charge collection efficiencies can be evaluated using the carrier mobility–lifetime ( follows :(1)I=I0μectrodes . The calrubidium . As a reeristics a of the equation :(2)R=Ip−ng light . The max formula :(3)LDR=2I0 is the amplitude of the current and τ is the time constant. The calculated rise time (rτ) and fall time (fτ) were 208.3 μs and 288.4 μs, respectively, which demonstrated the effective extraction of carriers at the interface between the perovskite and Au electrodes and the perovskite thin film’s high charge mobility. The quadruple-cation perovskite films’ ultra-fast response time and broad absorption spectrum from the ultraviolet to the near-infrared demonstrated their promising application as broadband photodetectors. A summary of the state-of-the-art research regarding perovskite photodetectors’ performance is presented in Moreover, response time is a key parameter that represents the speed of detecting transient light signals. Rise time is the time duration from 10% to 90% of the saturation current. As shown in −2 light illumination; this demonstrated good photosensitive stability under ambient conditions.The repeatability of photodetectors in detecting light is an important factor limiting their practical application. It is well known that detectors based on mixed-cation perovskite thin films suffer from severe baseline drift during continuous on–off due to ion migration and phase segregation. As shown in + proportion into the precursor solution helped obtain highly crystalline perovskite films with large grain sizes and few grain boundaries. A reduction in nonradiative or undesired recombination improved the carrier mobility–lifetime product (μτ). Hence, the photodetector based on mixed-cation perovskite (Rb0.05(CsMAFA)0.95) thin film exhibited a good linear response over a light intensity range of 0.5 mW cm−2 to 20 mW cm−2 with a maximum responsivity of 11.8 mA W−1 and an excellent on–off ratio of up to 7252. Simultaneously, the photodetector showed a broad absorption spectrum between 400 and 765 nm, with a maximum D* of 5.33 × 1011 Jones. The effective extraction of carriers at the interface between the perovskite and Au electrodes facilitated the device’s ultra-fast response time; the rτ and the fτ were 208.3 μs and 288.4 μs, respectively, with no significant decrease in photocurrent during multiple cycle measurements. These results verify perovskite’s potential as a next-generation high-performance photodetector.In conclusion, we developed a photodetector based on a mixed-cation perovskite thin film. The incorporation of an optimal Rb


Effects of hot pixels on pixel performance in light and dark environments have been investigated in pinned photodiode 0.18 μm backside illuminated CMOS image sensors irradiated by 10 MeV protons. After exposure to protons, hot pixels and normal pixels are selected from the whole pixel array, and their influences on key parameters are analyzed. Experimental results show that radiation-induced hot pixels have a significant impact on pixel performance in dark environments, such as dark signal nonuniformity, long integration time, and random telegraph signal. Hot pixels are caused by defects with complex structures, i.e., cluster defects. Furthermore, the dark current activation energy result confirms that the defects causing the hot pixels have defect energy levels close to mid-gap. Backside illuminated CMOS image sensors (BSI CIS) feature a higher sensitivity compared to the frontside illuminated CIS, and have been extensively used for space imaging missions ,2,3. TheMuch effort has been made to study displacement damage-induced hot pixels ,8,9,10, t damage ; (c) thet damage . Although these mechanisms have been proposed in previous work, it can be known that the generation of hot pixels depends on many factors, such as the electric field in the pixel, the defect types and density, and the defect energy level within the Si bandgap. Hence, displacement damage-induced hot pixel is still a complicated issue, and the dominant mechanism needs to be identified. Moreover, the previously published papers mainly focus on the behaviors of hot pixels. Additional work is needed to investigate the effects of hot pixels on pixel performance in light and dark environments. The purpose of this paper is to investigate the effects of hot pixels on pixel performance in light and dark environments and to provide a new perspective for understanding hot pixels. We randomly select the hot pixels and normal pixels from the whole array, and study their influences on key parameters, including full well capacity, quantum efficiency (QE), conversion gain (CVG), dark signal nonuniformity, and random telegraph signal (RTS). From the perspective of the defects, the correlation between the hot pixels and RTS pixels is discussed in detail. In addition, the physical mechanism of radiation-induced hot pixels is identified through dark current activation energy measurement.−, and the full well capacity is 90 ke−. The devices under study are the BSI CIS manufactured in a 0.18 μm CMOS process dedicated to imaging. The image sensors are constituted of 2048 × 2048-11 μm-pitch-4T pixels using the pinned photodiode (PPD). The image sensor supports two modes: STD mode at 48 fps and HDR mode, which is designed to achieve a high dynamic range (93 dB) at 24 fps. The HDR mode is selected to acquire raw images in this work. The shutter type of the image sensor is an electronic rolling shutter. The temporal dark current is 1.6 e9 p·cm−2, and the irradiation duration is 100 s. The cumulative displacement damage dose is 39.4 TeV/g, and the total ionizing dose is 2.8 krad(Si). The proton flux is 5 × 107 p·cm−2·s−1, and the proton beam homogeneity is within ±5%. The image sensors were irradiated at room temperature with all the pins grounded.BSI CIS was irradiated by 10 MeV protons at the Institute of Heavy Oon Physics, Peking University, CHN. The proton fluence is 5 × 10Key parameters test and characterization were performed at 24 °C one week after exposure to protons according to the standard for characterization of image sensors and cameras (EMVA1288). RTS pixel test was carried out with the successive acquisition of the raw images. The duration of raw image acquisition is 3 h, and the integration time is set to 1 s. RTS pixels can be detected from the subarray of the interest by an automated RTS test software, which is implemented using MATLAB software and is based on the sharp edge detection technique described in ref. . The RTSdark), which is independent of the particle types and energy [d energy ,17. Thus−/s, and the one is 415.4 e−/s for normal pixels). In order to obtain the response of the selected pixels in light and dark environments, the locations of the selected hot pixels and normal pixels are recorded to track these pixels.Hot pixels are located at the exponential tail, and normal pixels are in the Gaussian peak. According to the definition of the hot pixels, we select 10 hot pixels with the maximum dark current from the exponential tail and 10 normal pixels from the Gaussian peak. The normal pixel is randomly selected from the whole array and meets statistical requirements. 5 e−/s (corresponding to the photo-generated current). When the integration time is 0.2 s, the pixel output reaches full well capacity. Full well capacity is defined as the maximum amount of electrons stored in the pixel and is related to the light flux, VLOTG, and temperature [perature . The stuηλ) and can be converted into an electron (e−) through the CVG. CVG is defined as how much digital number variations are produced by an electron. − in the studied BSI CIS, and there is no difference in CVG for the hot pixels and normal pixels. Therefore, it can be concluded that radiation-induced hot pixels have no influence on the optoelectric performance in BSI CIS.QE represents the pixel’s ability to convert the incident photons to electrons at distinct wavelengths and is expressed as follows:−. Dark signal nonuniformity refers to dark signal variations from pixel to pixel, and its unit is eExcept for the hot pixel, the RTS pixel is another anomalous pixel in solid-state imaging devices and refers to dark current fluctuation between two or more discrete levels ,22. RTS dcI is the dark current, A is the preexponential factor, aE is the activation energy, k is the Boltzmann constant, and T is the temperature. Considering the temperature dependence of each parameter [tE is the defect energy level, iE is the mid-gap energy level, and gE is the Si bandgap. Considering the temperature dependence of the Si bandgap, we obtain the expression of activation energy as a function of the energy level into the silicon bandgap as:To harvest the energy level of the defects, dark current was tested at distinct temperatures, and the corresponding activation energy was calculated according to the Arrhenius law ,26,27. that the defect energy level is located at the mid-gap when the dark current activation energy is 0.649 eV, and the generation ability of the defect is maximized. In this paper, the effects of hot pixels on the pixel performance are investigated in pinned photodiode 0.18 μm backside illuminated CMOS image sensors irradiated by 10 MeV protons up to 39.4 TeV/g. After proton irradiation, hot pixels are produced by displacement damage through the elastic and inelastic nuclear scattering. Hot pixels are selected from the exponential tail of the dark current distribution, and normal pixels are randomly selected from the whole array. The method of selecting two kinds of pixels meets statistical requirements.Hot pixel has no influence on optoelectric performance since the photo-generated current is much larger than the dark current, and the impact of the dark current can be neglected. However, it is worth pointing out that the effects of hot pixels on pixel performance in dark environments should be a concern. Hot pixel behaves as a “bright spot” in the captured dark images and gives rise to the increase of dark signal nonuniformity, which has an impact on the dim target detection mission. Dark signal nonuniformity degradation is strongly dependent on the number of hot pixels. A hot pixel is able to reach the situation of full well capacity at the long integration time without light illumination. Above the specific integration time, the hot pixel makes no response and is not an effective pixel anymore. This case has an impact on the astronomical observation mission, in which the observation at the long integration time is required for the image sensors. It is confirmed that there is a correlation between hot pixels and RTS pixels, which is the hot pixels tend to exhibit RTS characteristics. The possible explanation is that the defects responsible for the hot pixel have a complex structure. The complex defects not only spontaneously transition from one configuration to another configuration due to the instability of the defects, but also have a big probability of introducing energy levels close to mid-gap. According to the characteristics of displacement damage-induced defects, cluster defects, which have complex defect structures, are more likely to cause the production of hot pixels. Furthermore, the result of dark current activation energy suggests that radiation-induced hot pixel is caused by defects with energy levels close to mid-gap.


Numerous studies have demonstrated that hepatic fibrosis, a progressive condition as an endpoint of multiple chronic hepatic diseases, is largely characterized with the extensive activation of hepatic stellate cells (HSCs). The precise effect of miR-488-5p in HSCs during hepatic fibrosis has not been elucidated.4), high-fat diet (HFD) and bile duct ligation (BDL). In vitro, the effects of miR-488-5p in HSCs were examined through cell proliferation assay and apoptosis. Luciferase reporter assay was applied to identify the underlying target of miR-488-5p. In vivo, the effects of miR-488-5p were explored through mouse liver fibrosis models.In our study, qRT‐PCR was applied to assess the level of miR-488-5p in activated HSCs stimulated by TGF-β1. We built murine liver fibrosis models with carbon tetrachloride  expression via straightly binding onto the 3′ UTR of its mRNA, which sequentially restrained the TGF-β/Smad2/3 pathway. TET3 inhibition induced by the overexpression of miR-488-5p reduced extracellular matrix deposition, which contributed to mitigating mouse liver fibrosis.We highlight that miR-488-5p restrains the activation of HSCs and hepatic fibrosis via targeting TET3 which is involved in the TGF-β/Smad2/3 signaling pathway. Collectively, miR-488-5p is identified as a potential therapeutic target for hepatic fibrosis.The online version contains supplementary material available at 10.1007/s12072-022-10404-w. However, under the effect of the longtime harmful factors, liver fibrosis will sequentially transform into liver cirrhosis, hypohepatia and even hepatocellular carcinoma [Hepatic fibrosis, as a self-repair mechanism, is quite common when stimuli such as alcohol, hepatitis B virus (HBV), carbon tetrachloride , belonging to a class of small non‐coding RNAs, serve as post-transcriptional regulators by negatively regulating downstream target gene expression , 10. Inc4, high-fat diet (HFD) and bile duct ligation (BDL) would be mitigatory once the mice were treated with ago-miR-488-5p. To sum up, our research findings demonstrate that miR-488-5p acts as a vital role in the hepatic fibrosis and might be a possible therapeutic target.The purpose of our research was intended to detect the role of miR-488-5p in hepatic fibrosis. First, the data presented that miR-488-5p were remarkably downregulated in the activated HSCs stimulated by TGF-β1 and mouse liver fibrosis models. Degree of activation of HSCs was restrained by miR-488-5p overexpression, as evidenced by the inhibited proliferation, promoted apoptosis and the reduction of profibrotic marker expression. We identified that tet methylcytosine dioxygenase 3 (TET3), serving as a target gene of miR-488-5p, played a pivotal role in hepatic fibrosis. More importantly, the degree of hepatic fibrosis attributed to CClAll fibrotic specimens from patients with advanced liver disease and normal tissues from patients with hepatic hemangioma were collected in the first Affiliated Hospital of Nanjing Medical University. All samples were straightly frozen in liquid nitrogen once the surgical resection was completed.We purchased the LX-2 cells, as a kind of cell line of hepatic stellate cells (HSCs), from the Cell Center of Shanghai Institutes for Biological Science. LX-2 cells were cultured with the Dulbecco’s Modified Eagle Medium  which contained 10% fetal bovine serum (FBS) and 1% antibiotics . Addition of TGF-β1 (10 ng/mL) into DMEM with no serum was intended to activate the LX-2 cells for 0, 3, 6, 12, and 24 h.4  through intraperitoneally injection twice a week for 8 weeks was intended to establish the hepatic fibrosis model. Meanwhile, olive oil (2 mL/kg) was used by the same way as the control group. The high-fat diet  was used to feed the mice for 24 weeks to establish the HFD model. Meanwhile, normal diet was used in the control group. For the control and bile duct ligation (BDL) group, sham or BDL surgery was performed in the mice, respectively.Male C57BL/6 mice, 8 weeks old, were obtained and fed based on standard conditions at the animal house of Nanjing Medical University. Treatment with CClThe RIPA buffer embracing 1% protease inhibitor and 1% PMSF was employed to extract the total protein based the standard protocol from the manufacturer (Beyotime). The extracted samples were separated via 10% SDS-PAGE and sequentially transferred to PVDF membranes. All antibodies were used in this study: TET3, TIMP-1 (Abcam), TGF-β, collagen-I, α-SMA, Smad2, p-Smad2, p-Smad3, Smad3, and GAPDH rabbit antibodies and HRP-conjugated anti-rabbit IgG antibodies .The TRIzol reagent was employed to extract total RNA on the basis of the standard instructions. Subsequently, RNA reversion and cDNA synthesis were carried out through using Reverse Transcription Kit (Vazyme). mRNA amplification and cDNA quantification were carried out through using SYBR green (Vazyme) on the basis of standard protocols. The levels of miR-488-5p and U6 were assessed via using TaqMan miRNA assay system (Life Technologies Corporation). The normalization of genetic level rests with U6 or β-actin level, respectively. The primers used are presented in Supplementary material.4 and HFD liver fibrosis models for 2 weeks was finished, we administrated ago-miR control and ago-miR-488-5p into the mice twice a week. After the treatment with ago-miR control and ago-miR-488-5p twice a week for 2 weeks, the liver tissues from BDL model mice were obtained. Eventually, we collected the liver tissues for further detection.miR-488-5p mimics, scrambled miRNA (SCR-miRNA), TET3 lentivirus (LV-TET3), and control lentivirus (LV-NC), ago-miR-488-5p and ago-miR control were obtained from GenePharma. Transfection process was auxiliary with the existence of lipofectamine 2000 (Invitrogen) for 48 h for further researches. Mice were subjected to the treatment with ago-miR control and ago-miR-488-5p (20 nmol/200 mL) through tail injection. Once establishing the CClWe planted LX-2 cells into 6-well plates at a density of 1000 cells per well and cultured for 15 days. The supernatant was discarded, and the clone was fixed with 4% paraformaldehyde. Staining was performed using a solution of crystal violet (staining time: 30 min). Eventually, the number of clone formations was counted.We planted LX-2 cells into 96-well plates at a density of 2000 cells per well and cultured them with DMEM supplemented with CCK-8 reagent (10 μL/well) for 5 days to evaluate the cell viability. Once incubation for 2 h was completed, the absorbance (450 nm) of each well was observed.The apoptotic level of LX-2 cells was evaluated through the Apoptosis Detection Kit . After incubation with 5 μL Annexin V-FITC and 5 μL PI for 20 min, each cell sample was mixed with 400 μL binding buffer. All steps were followed by manufacturer’s protocol.After liver tissues were totally fixed with 4% paraformaldehyde, they were embedded in paraffin. Subsequently, the paraffin block was made into slice and dyed with hematoxylin and eosin (HE). The Masson’s-stained or Sirius red’s-stained sections were used to assess the degree of liver fibrosis using the METAVIR scoring method. Eventually, the positive area was evaluated by pathologist to present the collagen deposition.After we obtained the tissue samples, the paraffin embedding was completed followed by standard steps. The paraffin block was made into slices (4 μm) which were incubated with anti-TET3 and anti-α-SMA for 12 h at 4 °C. Afterwards, the secondary antibody conjugated with HRP was utilized to incubate the slices for 1 h and eventually were dyed with 3,3′-diaminobenzidine and hematoxylin.All steps obeyed to the standard protocols of IF staining. In short, LX-2 cells were incubated with primary antibody α-SMA, and followed by secondary goat anti-rabbit Texas Red-conjugated IgG. At the same time, DAPI was used to present the nuclei.The luciferase reporter vector embracing the wild-type sequences of 3′‐UTR of TET3, and the mutated vector was constructed based on the binding sites. The luciferase reporter vectors were then co-transfected with miR‐488‐5p mimic or miR-SCR. At 48 h after transfection, cells were harvested and tested with the Dual-Luciferase reporter assay system  followed with manufacturer’s protocol. The relative luciferase activity was assessed as the ratio between firefly and Renilla luciferase activities detected by a dual-luciferase system .p value. Once p value < 0.05, was considered statistically significant.Statistical analysis results were shown as mean ± SEM. The SPSS software and GraphPad Prism were employed to manage the data and calculate the First, the LX-2 cells were cultured with medium containing TGF-β1 (10 ng/mL) for 0, 3, 6, 12 and 24 h. As shown in the Fig. 4, BDL and HFD were administrated to the mice to, respectively, develop liver fibrosis. As shown in the Fig. 4, BDL and HFD. Compared with the respective control (CTL) group, the histological structure was seriously damaged in CCl4 and BDL group, meanwhile the lipid vacuoles were abundant in the HFD group. At the same time the massive deposition of collagen was presented in CCl4, HFD and BDL group. Compared to their respective CTL group, the mRNA level of fibrosis-related markers containing α-SMA, tissue inhibitor of metalloproteinases 1 (TIMP-1) and Collagen-I were significantly upregulated  was a probable target gene of miR-488-5p. After consulting plentiful literatures, we found that tet methylcytosine dioxygenase 3 (TET3) was closely related with fibrosis , 15. ForFew studies have reported that TET3 played an essential role in activating TGF-β pathway, which is closely involved in the liver fibrosis , 16. For4, HFD and BDL group were, respectively, subjected to the transfection with ago-miR control (CTL) and ago-miR-488-5p. As presented in the Fig. 4 and BDL group; meanwhile, the number of lipid vacuoles was reduced in the HFD group   Fig. d. EventuHepatic fibrosis, as a common pathological condition in clinic, is usually caused by the long-term exposure to viral hepatitis, excessive drinking, fat-rich diet, drug-induced liver injury and biliary obstruction, and so on –19. One The role of miRNAs in HSCs activation and hepatic fibrosis has attracted plenty of attention for the past few years , 23. For4, BDL and HFD models and human fibrotic liver samples. Therefore, our study indicated that miR-488-5p might play a vital role in inhibiting hepatic fibrosis, which was worthy of further study.Through referring many literatures concerning miRNA and liver fibrosis, we noticed that the microarray analysis for miRNA expression from a previous study revealed miR-488-5p was significantly downregulated in activated LX-2 cells which hinted that miR-488-5p may be involved with the progression of liver fibrosis . Our datBioinformatics analyses were executed to explore the underlying target of miR-488-5p through miRWalk and TargetScan, the further experiments indicated that tet methylcytosine dioxygenase 3 (TET3) was the direct target of miR-488-5p in LX-2 cells. TET3 belongs to the member of ten–eleven translocation (TET) family proteins that participate in oxidizing 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC) . It had As we know, TGF-β participates in the plenty of biological processes, containing cellular migration, development, proliferation and differentiation, and so on . MoreoveIn conclusion, we identified that miR-488-5p, as a fibrosis-inhibitory factor, was downregulated in the activated HSCs and fibrotic liver tissues. miR-488-5p overexpression restrained HSCs activation and hepatic fibrosis via restraining TET3/TGF-β/SMAD2/3 signaling pathway.Supplementary file1 (DOCX 16 KB)Below is the link to the electronic supplementary material.