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The regulation of bile acid pathways has become a particularly promising therapeutic strategy for a variety of metabolic disorders, cancers, and diseases. However, the hydrophobicity of bile acids has been an obstacle to clinical efficacy due to off-target effects from rapid drug absorption. In this report, we explored a novel strategy to design new structure fragments based on lithocholic acid (LCA) with improved hydrophilicity by introducing a polar “oxygen atom” into the side chain of LCA, then (i) either retaining the carboxylic acid group or replacing the carboxylic acid group with (ii) a diol group or (iii) a vinyl group. These novel fragments were evaluated using luciferase-based reporter assays and the MTS assay. Compared to LCA, the result revealed that the two lead compounds 1a–1b were well tolerated in vitro, maintaining similar potency and efficacy to LCA. The MTS assay results indicated that cell viability was not affected by dose dependence (under 25 µM). Additionally, computational model analysis demonstrated that compounds 1a–1b formed more extensive hydrogen bond networks with Takeda G protein-coupled receptor 5 (TGR5) than LCA. This strategy displayed a potential approach to explore the development of novel endogenous bile acids fragments. Further evaluation on the biological activities of the two lead compounds is ongoing.

Concurrent translocations of MYC and BCL2 lead to abnormal expression of both oncoproteins, which contribute to the aggressive clinical characteristics of double-hit lymphoma (DHL). An effective therapy for DHL remains an unmet clinical need. In this study, we showed that both Ca

Chronic bacterial prostatitis (CBP) occurs frequently in the male population and significantly influences quality of life. Antibiotics are the main strategy for managing chronic bacterial prostatitis; however, most antibiotics have low efficacy due to their poor penetration of prostate tissues. To overcome this challenge, we fabricated cefpodoxime proxetil (CPD)-loaded reactive oxygen species (ROS)-responsive nanoparticles (NPs) for targeted treatment of CBP. These NPs were modified with folic acid (FA) and could be effectively internalized by bacteria-infected macrophages and prostatic epithelial cells because of the high expression of folate receptors (FRs) in these cells. In vitro cellular assays demonstrated that the CPD-loaded nanomedicine can obviously reduce proinflammatory cytokine expression in cells since the nanomedicine can efficiently eradicate cellular bacteria. In vivo imaging results verified that FA-modified nanomedicines can penetrate the prostatic epithelium and accumulate in the glandular lumen because FRs overexpression was also observed in the prostate tissues of CBP mice. Animal experiments demonstrated that FA-modified nanomedicine can remarkably relieve pelvic pain in CBP mice and dramatically decrease proinflammatory cytokine expression in prostate tissues via eradication of bacteria and scavenging of ROS. Our results provide a new strategy to deliver antibiotics for targeted therapy of CBP. STATEMENT OF SIGNIFICANCE: To overcome poor penetration of antibiotics in prostatic tissues, we developed an antibiotics-loaded ROS-responsive NPs for targeted treatment of CBP. We demonstrated that both bacteria-infected macrophages and prostatic epithelial cells have FRs overexpression, thus FA-modified NPs can be efficiently internalized by these cells. FA-modified NPs can penetrate the prostatic epithelium and accumulate in the glandular lumen via FRs-mediated endocytosis, and the accumulated NPs can smartly release their payload under high ROS microenvironment. A distinguished therapy outcome was obtained in murine CBP model since CPD-loaded NPs can efficiently eradicate the resident bacteria in prostate tissues and downregulate proinflammatory cytokine expression. Our work provides a practicable strategy to expand the application of antibiotics for management of CBP.

Background & Aims: Alcohol-associated liver disease (AALD) is one of the most common causes of liver injury and failure. Limited knowledge of the mechanisms underlying AALD impedes the development of efficacious therapies. Bile acid (BA) signaling was shown to participate in the progression of AALD. However, the mechanisms remain poorly understood. Methods: C57BL/6J wild-type (WT), Takeda G-protein–coupled bile acid receptor 5 (TGR5) knockout (KO) and brown adipose tissue (BAT)-specific TGR5 knockdown mice were subjected to ethanol feeding–induced AALD. Liver samples from alcoholic hepatitis patients were used to examine the BA circulation signaling. Human Embryonic Kidney Cells 293 were used for the TGR5 reporter assay. 23(S)-methyl-lithocholic acid was used as a molecular tool to confirm the regulatory functions of BAT in the AALD mouse model. Results: Ethanol feeding increased the expression of the thermogenesis genes downstream of TGR5 in BAT of WT, but not TGR5 KO, mice. TGR5 deficiency significantly blocked BAT activity and energy expenditure in mice after ethanol feeding. Alcohol increased serum BA levels in mice and human beings through altering BA transportation, and the altered BAs activated TGR5 signaling to regulate metabolism. Compared with ethanol-fed WT mice, ethanol-fed TGR5 KO mice showed less free fatty acid (FFA) β-oxidation in BAT, leading to higher levels of FFA in the circulation, increased liver uptake of FFAs, and exacerbated AALD. BAT-specific TGR5 knockdown mice showed similar results with TGR5 KO mice in AALD. Agonist treatment significantly activated TGR5 signaling in BAT, increased thermogenesis, reduced serum FFA level, and ameliorated hepatic steatosis and injury in AALD mice, while these effects were lost in TGR5 KO mice. Conclusions: BA signaling plays a protective role in AALD by enhancing BAT thermogenesis. Targeting TGR5 in BAT may be a promising approach for the treatment of AALD.

The Myc proto-oncogene family consists of three members, C-MYC, MYCN, and MYCL, which encodes the transcription factor c-Myc (hereafter Myc), N-Myc, and L-Myc, respectively. Myc protein orchestrates diverse physiological processes, including cell proliferation, differentiation, survival, and apoptosis. Myc modulates about 15% of the global transcriptome, and its deregulation rewires the cellular signaling modules inside tumor cells, thereby acquiring selective advantages. The deregulation of Myc occurs in >70% of human cancers, and is related to poor prognosis; hence, hyperactivated Myc oncoprotein has been proposed as an ideal drug target for decades. Nevertheless, no specific drug is currently available to directly target Myc, mainly because of its “undruggable” properties: lack of enzymatic pocket for conventional small molecules to bind; inaccessibility for antibody due to the predominant nucleus localization of Myc. Although the topic of targeting Myc has actively been reviewed in the past decades, exciting new progresses in this field keep emerging. In this review, after a comprehensive summarization of valuable sources for potential druggable targets of Myc-driven cancer, we also peer into the promising future of utilizing macropinocytosis to deliver peptides like Omomyc or antibody agents to intracellular compartment for cancer treatment.

Background Oxygen plays a pivotal role in cardiopulmonary resuscitation (CPR) and postresuscitation intervention for cardiac arrest. However, the optimal method to reoxygenate patients has not been determined. This study investigated the effect of timing of hyperoxygenation on neurological outcomes in cardiac arrest/CPR rats treated with targeted temperature management. Methods and Results After induction of ventricular fibrillation, male Sprague‐Dawley rats were randomized into 4 groups (n=16/group): (1) normoxic control; (2) O 2 _CPR, ventilated with 100% O 2 during CPR; (3) O 2 _CPR+postresuscitation, ventilated with 100% O 2 during CPR and the first 3 hours of postresuscitation; and (4) O 2 _postresuscitation, ventilated with 100% O 2 during the first 3 hours of postresuscitation. Targeted temperature management was induced immediately after resuscitation and maintained for 3 hours in all animals. Postresuscitation hemodynamics, neurological recovery, and pathological analysis were assessed. Brain tissues of additional rats undergoing the same experimental procedure were harvested for ELISA‐based quantification assays of oxidative stress–related biomarkers and compared with the sham‐operated rats (n=6/group). We found that postresuscitation mean arterial pressure and quantitative electroencephalogram activity were significantly increased, whereas astroglial protein S100B, degenerated neurons, oxidative stress–related biomarkers, and neurologic deficit scores were significantly reduced in the O 2 _CPR+postresuscitation group compared with the normoxic control group. In addition, 96‐hour survival rates were significantly improved in all of the hyperoxygenation groups. Conclusions In this cardiac arrest/CPR rat model, hyperoxygenation coupled with targeted temperature management attenuates ischemia/reperfusion‐induced injuries and improves survival rates. The beneficial effects of high‐concentration oxygen are timing and duration dependent. Hyperoxygenation commenced with CPR, which improves outcomes when administered during hypothermia.

Additional file 7: Supplementary Fig. 4. ATAD3A is down-regulated in hypoxia and a potential target of miR-210-5P. a Immunofluorescent staining of HIF-1α (green) and ATAD3A (red) expression in normoxic and hypoxic LM3 cells (1% O2 treatment for 24 h). Scale bar = 200 μm. b Immunoblot showing expression levels of indicated proteins in Huh7 cells with/without CoCl2 treatment for different durations. c qRT-PCR analysis of ATAD3A expression in LM3 cells with/without CoCl2 treatment. d Bioinformatics analysis of the hypoxia-responsive miRNAs from digestive cancers. e qRT-PCR analysis showing miR-210-5P levels in Huh7 and LM3 cells treated with DMSO or CoCl2 for 24 h. f qRT-PCR analysis showing the expression of ATAD3A in LM3 cells treated with miR-210-5P mimic and inhibitor. Each bar represents the mean ± SEM of three independent experiments. *P

Additional file 2. Antibodies used for immunostaining analysis in our dataset

Additional file 6: Supplementary Fig. 3. ATAD3A plays a vital role in lactate production of hepatoma cells. a,b Lactate levels in the supernatants of Huh7-shControl and Huh7-shATAD3A (a) or LM3- shControl and LM3-shATAD3A (b) cells. c,d Lactate levels in Huh7-Vector and Huh7-ATAD3A (c) or LM3-Vector and LM3-ATAD3A (d) cells with/without 1% O2 exposure for 24 h. e,f Lactate levels in Huh7 (e) and LM3 (f) cells with/ without miR-210-5P inhibitor transfection and with/ without CoCl2 treatment. g, h Lactate levels in control and miR-210-5P mimic-transfected Huh7 (g) and LM3 (h) cells. Each bar represents the mean ± SEM of three independent experiments. *P

Additional file 1. Target sequences and ORF expression clone for PINK1 and ATAD3A in the study.

Additional file 3. Sequences of the primers used for qRT-PCR.

Additional file 4: Supplementary Fig. 1. Hypoxia induced sorafenib resistance in hepatoma cells. a,b Sorafenib sensitivity under normoxic or hypoxia condition of LM3 and Huh7 cells measured by apoptosis rates (%). c, d Cell viability of LM3(c) or Huh7 (d) cells under hypoxic or normoxic conditions with sorafenib treatment. e qRT-PCR analysis of indicated genes in control, hypoxia treated, sorafenib resistant and hypoxia-induced sorafenib resistant Huh7 cells. f. Immunoblot showing expression levels of indicated proteins in control, hypoxia, SR and Huh7-H-SR cells. g-i Relation between HIF-1α expression and that of ABCC1 (g), TOMM20 (h) and TOMM70 (i) in HCC patients from TCGA database. Each bar represents the mean ± SEM of three independent experiments. *P

AE (alopecia and excoriation) is a mouse mutant phenotype that harbours a mutation in Gsdma3. Gsdma3 has been reported to regulate the development of skin and hair follicles. However, its role in the mammary glands has not been reported. In this study, we found that descendants bred from an AE mother died within 12 days after birth. Then, we found that the expression of Gsdma3 varied among the developmental stages of mammary glands. Subsequently, we systematically studied the phenotype of the mammary glands of AE and wild-type mice, revealing that the mammary glands were smaller in AE mice. The mammary glands of AE mice exhibited shorter ductal extension and less bifurcation. Immunohistochemistry staining indicated that the mammary glands of AE mice displayed more proliferating cells during puberty while secreting less β-casein during pregnancy and lactation. The lymph nodes in the mammary glands of the AE mice were larger and showed some pigmentation, suggesting that the immune reaction in the mammary glands was up-regulated. Under a transmission electron microscope, residual bodies were observed in the lymph nodes in the mammary glands of AE mice. Thus, we report a new function of Gsdma3 in regulating the development of mammary glands, and we demonstrate that the Gsdma3 gene may act as a suppressor of the immune reaction.

The impact between acetabulum and femoral neck is another possible mechanism of femoral neck fracture. Direct trauma of the greater trochanter may not be able to fully explain the mechanism underlying femoral neck fracture. In this study, we sought to investigate whether anterior femoroacetabular impingement are associated with femoral neck fractures. A total of 36 patients with femoral neck fracture who had undergone total hip arthroplasty or hemiarthroplasty were included in this study. These patients were divided into 2 groups: labrum tear group and normal labrum group. Patients’ age, gender, body mass index, muscle injury, injury pattern, trauma severity, femoral head-neck offset, femoral head-neck ratio, Cam deformity alpha angle, acetabular anteversion, femoral head diameter, acetabular index, cortical index, hip axis length, and neck stem angle were recorded and analyzed. SPSS 18.0 software was used for statistical analyses. According to intraoperative findings, 22 patients exhibited a labrum tear. Magnetic resonance imaging examination revealed bone contusion on the anterolateral margin of the acetabulum with muscle damage surrounding the hip. Among 14 cases without a labrum tear, no bone contusion and obvious muscle injury were found on the anterolateral margin of the acetabulum. Notably, muscle injury, injury pattern, trauma severity and femoral head-neck offset differed significantly (P

Hepatocellular carcinoma (HCC) is the most prevalent malignancy in liver and a leading cause of cancer-related deaths. Despite the pressing need for treatment options, patients with HCC develop significant resistance and adverse side effects to current approved drugs that becomes a major barrier to effective treatment. A natural product Tetrandrine (TET) is a potential alternative treatment option for HCC, with demonstrated effectiveness and low toxicity. However, the mechanisms by which Tetrandrine inhibits HCC are unclear. In the current study, we identify Ca

Adenosine A2A receptors (A2ARs) in distinct cellular types may exert different and even opposite effects on many neurologic disorders; A2ARs in bone marrow-derived cells (BMDCs) have been shown to play important roles in various brain injuries. We previously showed that global A2AR inactivation aggravates chronic cerebral hypoperfusion-induced white matter lesions (WMLs); however, the specific cell populations responsible for A2AR-mediated signaling remain unknown. In the present study, we developed chimeric mice in which A2ARs were either selectively inactivated or reconstituted in BMDCs by transplanting bone marrow from global A2AR gene knockout or wild-type mice into wild-type or gene knockout mice, respectively. Chimeric mice were subsequently subjected to chronic cerebral hypoperfusion by bilateral common carotid artery stenosis, and the effects of BMDC A2ARs on WMLs were evaluated. The selective inactivation of A2AR in BMDCs aggravated chronic cerebral hypoperfusion-induced WMLs, promoted microglial activation, and increased proinflammatory cytokine expression, whereas the selective reconstitution or activation of A2AR in BMDCs using the agonist CGS21680 produced the opposite effects. These results demonstrate that A2ARs in BMDCs are important modulators of WMLs induced by chronic cerebral hypoperfusion; this modulation might be associated with the regulation of inflammatory cytokine production.

Background & Aims Bile acid transporters maintain bile acid homeostasis. Little is known about the functions of some transporters in cholestasis or their regulatory mechanism. We investigated the hepatic expression of solute carrier organic anion transporter family member 3A1 (SLCO3A1, also called OATP3A1) and assessed its functions during development of cholestasis. Methods We measured levels of OATP3A1 protein and messenger RNA and localized the protein in liver tissues from 22 patients with cholestasis and 21 patients without cholestasis, using real-time quantitative polymerase chain reaction, immunoblot, and immunofluorescence analyses. We performed experiments with Slco3a1-knockout and C57BL/6J (control) mice. Mice and Sprague-Dawley rats underwent bile duct ligation (BDL) or a sham operation. Some mice were placed on a 1% cholic acid (CA) diet to induce cholestasis or on a control diet. Serum and liver tissues were collected and analyzed; hepatic levels of bile acids and 7-α-C4 were measured using liquid chromatography/mass spectrometry. Human primary hepatocytes and hepatoma (PLC/PRF/5) cell lines were used to study mechanisms that regulate OATP3A1 expression and transport. Results Hepatic levels of OATP3A1 messenger RNA and protein were significantly increased in liver tissues from patients with cholestasis and from rodents with BDL or 1% CA diet–induced cholestasis. Levels of fibroblast growth factor 19 (FGF19, FGF15 in rodents) were also increased in liver tissues from patients and rodents with cholestasis. FGF19 signaling activated the Sp1 transcription factor and nuclear factor κB to increase expression of OATP3A1 in hepatocytes; we found binding sites for these factors in the SLCO3A1 promoter. Slco3a1-knockout mice had shorter survival times and increased hepatic levels of bile acid, and they developed more liver injury after the 1% CA diet or BDL than control mice. In hepatoma cell lines, we found OATP3A1 to take prostaglandin E2 and thyroxine into cells and efflux bile acids. Conclusions We found levels of OATP3A1 to be increased in cholestatic liver tissues from patients and rodents compared with healthy liver tissues. We show that OATP3A1 functions as a bile acid efflux transporter that is up-regulated as an adaptive response to cholestasis.