A MDM2 inhibitor MX69 inhibits adipocytes adipogenesis and differentiation.

Adipose tissue, a key regulator of systemic energy homeostasis, can synthesize and store triglycerides to meet long-term energy demands. In response to nutrient overload, adipose tissue expands by hypertrophy or hyperplasia. As an oncogene, MDM2 has exerted diverse biological activities including human development, tissue regeneration, and inflammation, in addition to major oncogenic activities. Recently, some studies indicated that MDM2 plays an important role in adipose tissue function. However, the role of MX69, a MDM2 inhibitor, in adipose tissue function has not been fully elucidated. Here, we administered MX69 intraperitoneally to high-fat diet-induced obesity (DIO) wild type C57BL/6 mice and found that MX69 could promote the body weight and white adipose tissue weight of DIO mice. Moreover, MX69 had no effects on glucose tolerance and insulin sensitivity in DIO mice. And MX69 treatment decreased the size of adipocytes and fat deposition in adipose tissue and inhibited 3T3-L1 preadipocytes differentiation. Mechanistically, MX69 inhibited the protein levels of MDM2 and the mRNA levels of genes related to adipogenesis and differentiation. In summary, our results indicated that MDM2 has a crucial and complex role in regulating adipose tissue function.

A digitalized isothermal nucleic acid testing platform based on a pump-free open droplet array microfluidic chip.

Digital PCR has shown great potential for quantitative nucleic acid testing (NAT), but most existing platforms are dependent on large auxiliary equipment (e.g., vacuum pump, amplification instrument, fluorescence microscope) to achieve target dispersion, amplification, signal capture and result analysis. Such complex, expensive and bulky NAT platforms have limited their applications in resource-limited areas, especially for point-of-care testing (POCT). In this work, we designed a digital isothermal NAT platform based on a pump-free open droplet array microfluidic chip. A pump-free microfluidic chip was developed based on an open microdroplet array in the form of thousands of independent microdroplets for spontaneous sample dispersion, without the need for external power. Combined with a handheld fluorescent signal reader based on a smartphone, this digital NAT platform can accurately quantify as low as 1 copy per µL of λDNA. Therefore, our integrated NAT platform, as a potable, robust and low-cost tool for highly accurate NA quantitative analysis, holds great potential for POCT applications.

Spermidine ameliorates non-alcoholic fatty liver disease through regulating lipid metabolism via AMPK.

In this study, treatment of high-fat diet-induced obesity (DIO) C57BL/6J mice with spermidine decreased body weight and subcutaneous and visceral fat content, reversed the apparent hepatosteatosis, and reduced hepatic intracellular and serum triglyceride and total cholesterol concentrations. Moreover, spermidine treatment improved glucose tolerance and insulin sensitivity in DIO mice. The mechanism studies indicated that spermidine indeed increased the phosphorylation of hepatic AMP-activated protein kinase (AMPK), and inhibited the expression of lipogenic genes in vivo and in vitro. Moreover, these spermidine-mediated molecular effects were also abolished by compound C, an inhibitor of AMPK, in primary hepatocytes. In summary, spermidine protected against DIO-induced hepatosteatosis by decreasing lipogenic genes expression through an AMPK-mediated mechanism.

Dual-crosslinked bioadhesive hydrogel as NIR/pH stimulus-responsiveness platform for effectively accelerating wound healing.

Adhesive hydrogels have emerged as promising candidates to solve life-threatening infectious skin injuries. However, the inadequate mechanical characteristics and biological adherence limit the traditional wound dressing unable to adapt to high-frequency movement and real-time monitoring of wound healing, calling for the development of bioadhesive materials guided wound healing. In this work, a multifunctional bioadhesive hydrogel with double colorimetric-integrated of polyethylene glycol (PVA)-dextran (Dex)-borax-bromothymol blue (BTB)-fluorescein thiocyanate (FITC) and functionalization by tungsten disulfide-catechol nanozyme (CL/WS2) was created. Hydrogel is a perfect biological adhesive, which can achieve repeatable and strong tissue adhesion strength (8.3 ± 0.6 kPa), which is 1.66 times that of commercial dressings. Based on the strong biological adhesion of the hydrogel, a sensor is integrated into the hydrogel to collect visual image of bacterial infection from a smartphone and transform it into an on-site pH signal for remote evaluation of the wound's dynamic status in real time. Ultimately, the adhesiveness hydrogel has high worth in managing the burden related to wound healing and paving the way for intelligent wound management in the future.

NIR as a "trigger switch" for situ distinguish superbacteria and photothermal synergistic antibacterial treatment with Ag2O particles/lignosulfonate/cationic guar gum hybrid hydrogel.

The in situ identification of superbugs with the simultaneous killing of it is key to preventing human health. Here, a one-stop identification and killing platform for near-infrared (NIR) triggering was designed and constructed using lignosulfonate (LS), cationic guar gum (CG) and Ag2O NPs hydrogels (LS/CG/Ag2O). The hydrogel network is used as a fixed matrix for Ag2O NPs and a nano reactor, meanwhile 3,3', 5,5'-tetramethylbenzidine (TMB) as a single probe sensor array for bacterial identification. In contrast to conventional methods, hybrid hydrogels have catalytic qualities through which TMB be catalyzed to generate oxidized TMB (oxTMB). The drug resistance of the same strain can be distinguished based on the different inhibition abilities of drug-resistant superbacteria in TMB and hydrogel reactions. Then, the employing of oxTMB photothermal characteristics, it can be efficiently killed in real time while being driven by a near-infrared laser. The proposed one-stop hydrogel platform paves a way for the rapid identification and killing of drug-resistant superbacteria.

Protocol to knock in or out target genes in mouse epidydimal white adipose tissue by spot injecting adeno-associated viruses.

Visceral fat obesity is more strongly associated with ectopic fat deposition, lipotoxicity, and metabolic disease compared to generalized obesity. To study the function of visceral fat tissue, we describe steps to knock in or out target genes by spot injecting adeno-associated viruses (AAV) in visceral fat tissue. We provide details on anesthesia, incision, and spot injection into the epidydimal white adipose tissue (eWAT) of live anesthetized mice. Furthermore, we detail an efficient technique for expressing exogenous protein in mouse eWAT. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2022).

Murine double minute 2 aggravates adipose tissue dysfunction through ubiquitin-mediated six-transmembrane epithelial antigen of prostate 4 degradation.

Healthy adipose tissue is crucial to maintain normal energy homeostasis. Little is known about the role of murine double minute 2 (MDM2), an E3 ubiquitin ligase and has been highlighted in oncopathology, in adipose tissue. Our results indicated that MDM2 expression was associated with nutritional status. Mdm2 adipocyte-specific knock-in (Mdm2-AKI) mice exhibited exacerbated weight gain, insulin resistance, and decreased energy expenditure. Meanwhile, chronic high-fat diet (HFD) exposure caused obvious epididymal white adipose tissue (eWAT) dysfunction, such as senescence, apoptosis, and chronic inflammation, thereby leading to hepatic steatosis in Mdm2-AKI mice. Mechanically, MDM2 could interact with six-transmembrane epithelial antigen of prostate 4 (STEAP4) and inhibit STEAP4 expression through ubiquitin-mediated STEAP4 degradation. Thereinto, the K18 and K161 sites of STEAP4 were ubiquitin-modificated by MDM2. Finally, STEAP4 restoration in eWAT of Mdm2-AKI mice on a HFD rescued MDM2-induced adipose dysfunction, insulin resistance, and hepatic steatosis. Summary, the MDM2-STEAP4 axis in eWAT plays an important role in maintaining healthy adipose tissue function and improving hepatic steatosis.