Inhibition of KIF11 ameliorates osteoclastogenesis via regulating mTORC1-mediated NF-κB signaling.

Osteoporosis, characterized by over-production and activation of osteoclasts, has become a major health problem especially in elderly women. In our study, we first tested the effect of Caudatin (Cau) in osteoclastogenesis, which is separated from Cynanchum auriculatum as a species of C-21 steroidal glyosides. The results indicated that Cau suppressed osteoclastogenesis in a time- and dose-dependent manner in vitro. Mechanistically, Cau was identified to inhibit NF-κB signaling pathway via modulation of KIF11-mediated mTORC1 activity. In vivo, by establishing an ovariectomized (OVX) mouse model to mimic osteoporosis, we confirmed that Cau treatment prevented OVX-induced bone loss in mice. In conclusion, we demonstrated that Cau inhibited NF-κB signaling pathway via modulation of KIF11-mediated mTORC1 activity to suppress osteoclast differentiation in vitro as well as OVX-induced bone loss in vivo. This provides the possibility of a novel prospective drug for osteoporosis remedies.

Discovering the Secret of Diseases by Incorporated Tear Exosomes Analysis via Rapid-Isolation System: iTEARS.

Nanoscale small extracellular vesicles (sEVs, exosomes) in tears allow us to investigate the multisignatures of diseases. However, the translations of tear sEVs for biomarker discovery and clinical diagnostics are practically limited by low recovery, long processing time, and small sample volume. Here, we report an incorporated tear-exosomes analysis via rapid-isolation system (iTEARS) via nanotechnology to discover the secrets of ocular disorders and systemic diseases. We isolate exosomes rapidly with high yield and purity from a few teardrops (∼10 µL) within 5 min via nanoporous membrane-based resonators for the quantitative detection and biomarker discovery through proteomic and transcriptomic analysis. We have identified 904 proteins, among which 228 proteins are discovered, 426 proteins are detected from exosomes of dry eye disease, and demonstrate CALML5, KRT6A, and S100P for the classification of dry eye disease. We have also investigated 484 miRNAs in tear exosomes and show miR-145-5p, miR-214-3p, miR-218-5p, and miR-9-5p are dysregulated during diabetic retinopathy development. We believe iTEARS can be used for improving molecular diagnostics via tears to identify ocular disorders, systemic diseases, and numerous other neurodegenerative diseases and cancer.

The Association of Aspartate Aminotransferase/Alanine Aminotransferase Ratio with Diabetic Nephropathy in Patients with Type 2 Diabetes.

PURPOSE: To investigate the relationship between aspartate aminotransferase to alanine aminotransferase ratio (AST/ALT) and diabetic nephropathy (DN). PATIENTS AND METHODS: A total of 402 patients with type 2 diabetes mellitus were divided into three groups, such as normoalbuminuria (n = 196), microalbuminuria (n = 131) and macroalbuminuria (n = 75) groups. Basic information and laboratory results were collected. Serum AST/ALT, tumor necrosis factor-α (TNF-α), interleukin (IL)-2, IL-4, IL-6, IL-10 and interferon- γ (INF- γ) were also measured. DN was defined as microalbuminuria or macroalbuminuria. The estimated glomerular filtration rate (eGFR) was calculated using the following formula: 186 × (serum creatinine)-1.154× (age)-0.203× (0.742 if female). RESULTS: The AST/ALT in the macroalbuminuria group was higher than in the microalbuminuria and normoalbuminuria groups. The concentrations of tumor necrosis factor-α (TNF-α), IL-2, IL-4, IL-10 and INF-γ in the macroalbuminuria group were significantly higher than those in the two other groups. Multivariate logistical analysis showed that after adjusting confounding factors, TNF-α and high AST/ALT were independent risks for DN and macroalbuminuria. Furthermore, the AST/ALT had significantly positive correlation with TNF-α (r = 0.101, P = 0.048), IL-4 (r = 0.185, P = 0.005) and IL-6 (r = 0.274, P < 0.001) levels. CONCLUSION: This study showed that high AST/ALT was an independent risk factor for the DN. Additionally, AST/ALT was positively correlated with inflammation cytokines, such as TNF-α, IL-4 and IL-6 levels.

Morin attenuates osteoclast formation and function by suppressing the NF-κB, MAPK and calcium signalling pathways.

Morin is a natural compound isolated from moraceae family members and has been reported to possess a range of pharmacological activities. However, the effects of morin on bone-associated disorders and the potential mechanism remain unknown. In this study, we investigated the anti-osteoclastogenic effect of morin in vitro and the potential therapeutic effects on ovariectomy (OVX)-induced osteoporosis in vivo. In vitro, by using a bone marrow macrophage-derived osteoclast culture system, we determined that morin attenuated receptor activator of nuclear factor (NF)-κB ligand (RANKL)-induced osteoclast formation via the inhibition of the mitogen-activated protein kinase (MAPK), NF-κB and calcium pathways. In addition, the subsequent expression of nuclear factor of activated T cells c1 (NFATc1) and c-fos was significantly suppressed by morin. In addition, NFATc1 downregulation led to the reduced expression of osteoclastogenesis-related marker genes, such as V-ATPase-d2 and Integrin ß3. In vivo, results provided that morin could effectively attenuate OVX-induced bone loss in C57BL/6 mice. In conclusion, our results demonstrated that morin suppressed RANKL-induced osteoclastogenesis via the NF-κB, MAPK and calcium pathways, in addition, its function of preventing OVX-induced bone loss in vivo, which suggested that morin may be a potential therapeutic agent for postmenopausal osteoporosis treatment.

Low-dose adefovir dipivoxil-induced hypophosphatemia osteomalacia in five chronic hepatitis B virus-infected patients. Is low-dose adefovir dipivoxil-induced nephrotoxicity completely reversible?

Adefovir dipivoxil (ADV) is one of the most important nucleostide analogues currently in use for the treatment of chronic hepatitis B virus (HBV) infection. Low-dose ADV-induced nephrotoxicity in most cases was reported to be reversible after the discontinuation of ADV or by decreasing the dose of ADV. In our study, we have 5 documented cases of low-dose ADV-induced hypophosphatemia osteomalacia with or without Fanconi syndrome which were diagnosed in our hospital between 2010 and 2017. Three patients were observed to have a full recovery after the discontinuation of ADV. Two patients had persistently elevated urine ß2-microglobulin levels and out of these two patients, one patient had persistent hypophosphatemia after the cessation of ADV. These cases illustrated that the use of low-dose ADV increased the risk of nephrotoxicity, and in some patients, low-dose ADV-induced nephrotoxicity was not completely reversible. Patients of East Asian origin, especially those with a low body mass index, were prone to a relatively higher risk of developing low-dose ADV-induced nephrotoxicity; therefore, it was worth paying attention to the side effects caused by low-dose ADV.

[CpG-ODN activation of TLR9 signaling pathways and NF-κB in STZ-induced diabetic rats].

OBJECTIVE: To observe the mRNA and protein expression of TLR9, NF-κB and serum level of Th1/Th2 cytokines when stimulated with different concentration of CpG-ODN in STZ-induced diabetic rats. METHODS: 40 SD male rats were randomly divided into four groups:normal control group (Group I), type 1 diabetes control group (Group II), low-dose CpG-ODN treated type 1 diabetes group (Group III), high dose CpG-ODN treated type 1 diabetes group (Group IV). 60 µg /kg and 120 µg /kg CpG-ODN were injected to rats of Group III and IV when inducing type 1 diabetes model.mRNA and protein levels of TLR9 and NF-κB in spleen and pancreas were detected by fluorescence quantitative PCR and Western blotting.Serum TNF-α, IL-6 and IFN-γ were measured by ELISA. RESULTS: TLR9 protein expression levels were much higher in spleen than in the pancreas among these four groups. In spleen, TLR9 and NF-κB mRNA and protein level were significantly higher in Group III and IV than those in Group I and II, and the expression level was higher in Group IV than in Group III. In pancreas, TLR9 and NF-κB protein level were significantly higher in Group III and IV than those in Group I and II, and the expression level was also higher in Group IV than Group III.Serum TNF-α and IFN-γ level in Group III and IV significantly higher than in Group I and II, and serum levels of TNF-α and IFN-γ in Group IV was higher than in Group III. CONCLUSIONS: CpG-ODN activation of TLR9-MyD88 signaling pathways results in dose-effect NF-κB stimulation in STZ-induced diabetic rats.