Role of AOX1 on RXR signaling regulates osteoblastogenesis in hPDLMSCs.

Alveolar bone loss resulting from periodontal disease ultimately leads to tooth loss. Periodontal ligament mesenchymal stem cells (PDLMSCs) are the tissue-specific cells responsible for maintaining and repairing the periodontal ligament, cementum, and alveolar bone. In this study, we explored the role of aldehyde oxidase 1 (AOX1) in regulating the osteoinduction of human periodontal ligament stem cells (hPDLMSCs). hPDLMSCs were isolated from clinically healthy donors, and AOX1 expression was assessed by comparing inducted and non-inducted hPDLMSCs. Remarkably, we observed a significant upregulation of AOX1 expression during osteoinduction, while AOX1 silencing resulted in the enhanced osteogenic potential of hPDLMSCs. Subsequent experiments and analysis unveiled the involvement of retinoid X receptor (RXR) signaling in the inhibition of osteogenesis in hPDLMSCs. Ligands targeting the RXR receptor mirrored the effects of AOX1 on osteogenesis, as evidenced by alterations in alkaline phosphatase (ALP) activity and bone formation levels. Collectively, these findings underscore the potential regulatory role of AOX1 via RXR signaling in the osteogenesis of hPDLMSCs. This elucidation is pivotal for advancing hPDLMSC-based periodontal regeneration strategies and lays the groundwork for the development of targeted therapeutic interventions aimed at enhancing bone formation in the context of periodontal disease.

Development and validation of nomograms to predict survival of neuroendocrine carcinoma in genitourinary system: A population-based retrospective study.

Neuroendocrine carcinoma (NEC) is a rare yet potentially perilous neoplasm. The objective of this study was to develop prognostic models for the survival of NEC patients in the genitourinary system and subsequently validate these models. A total of 7125 neuroendocrine neoplasm (NEN) patients were extracted. Comparison of survival in patients with different types of NEN before and after propensity score-matching (PSM). A total of 3057 patients with NEC, whose information was complete, were extracted. The NEC influencing factors were chosen through the utilization of the least absolute shrinkage and selection operator regression model (LASSO) and the Fine & Gary model (FGM). Furthermore, nomograms were built. To validate the accuracy of the prediction, the efficiency was verified using bootstrap self-sampling techniques and receiver operating characteristic curves. LASSO and FGM were utilized to construct three models. Confirmation of validation was achieved by conducting analyses of the area under the curve and decision curve. Moreover, the FGS (DSS analysis using FGM) model produced higher net benefits. To maximize the advantages for patients, the FGS model disregarded the influence of additional occurrences. Patients are expected to experience advantages in terms of treatment options and survival assessment through the utilization of these models.

Development and validation of nomograms to predict survival of primary cervical lymphoma: A surveillance, epidemiology, and end results (SEER) database analysis.

Due to the rarity of primary cervical lymphoma (PCL), the long-term survival of patients with cervical lymphoma and factors influencing survival are unknown. This study aimed to compare the survivals of patients with PCL and those with other cervical tumors and construct a clinical prediction model to assess the prognosis of patients with PCL. Patients with PCL from the Surveillance, Epidemiology, and End Results database were allocated randomly in a 7:3 ratio to the training and validation sets. Cox proportional hazard and Fine-Gray models were used to verify independent factors influencing overall survival (OS) and disease-specific survival (DSS), and nomograms were constructed. Receiver operating characteristic curve analysis and decision curve analysis (DCA) were used to test the performance and clinical utility of the models, respectively. We included 206 patients with PCL. The areas under the curves (AUCs) and DCA showed that all models had clinical benefits; The models constructed in this study had a predictive performance for patients with PCL. It can guide clinicians to rationalize the treatment plan for patients.

The association between three prevalent autoimmune disorders and the likelihood of developing prostate cancer: a Mendelian randomization study.

Numerous studies establish a significant correlation between autoimmune disorders (AIDs) and prostate cancer (PCa). Our Mendelian randomization (MR) analysis investigates the potential connection between rheumatoid arthritis (RA) and PCa, aiming to confirm causal links between systemic lupus erythematosus (SLE), hyperthyroidism, and PCa. Summary statistics from genome-wide association studies provided data on PCa and three AIDs. MR analysis, using IVW as the main approach, assessed causal relationships, validated by sensitivity analysis. IVW revealed a correlation between genetically anticipated RA and PCa, notably in Europeans (OR = 1.03; 95% CI 1.01-1.04, p = 2*10-5). Evidence supported a lower PCa risk in individuals with SLE (OR = 0.94; 95% CI 0.91-0.97, p = 2*10-4) and hyperthyroidism (OR = 0.02; 95% CI 0.001-0.2, p = 2*10-3). Weighted mode and median confirmed these findings. No pleiotropic effects were observed, and MR heterogeneity tests indicated dataset homogeneity. Our study establishes a causal link between RA, SLE, hyperthyroidism, and PCa.

Renal pelvis metastasis following surgery for breast angiosarcoma: a case report and literature review.

Renal metastasis of breast angiosarcoma is rare. This article reports the medical records of a patient diagnosed with breast angiosarcoma who underwent radical mastectomy and was found to have multiple lung metastases 3 years after surgery and renal pelvic metastasis 4 years after surgery. The patient underwent robot-assisted laparoscopic radical nephroureterectomy and sleeve resection of the intramural segment of the ureter, and postoperative pathology and immunohistochemical staining confirmed the diagnosis of renal pelvic metastasis of breast angiosarcoma. The patient received anlotinib for lung metastases following surgery and was followed up for 4 months after surgery. Currently, the patient has symptoms of coughing and hemoptysis but no other discomfort. The diagnosis and treatment of this rare malignant tumor remain challenging.

Protein-centric omics analysis reveals circulating complements linked to non-viral liver diseases as potential therapeutic targets.

BACKGROUND/AIMS: To evaluate the causal correlation between complement components and non-viral liver diseases and their potential use as druggable targets. METHODS: We conducted Mendelian randomization (MR) to assess the causal role of circulating complements in the risk of non-viral liver diseases. A complement-centric protein interaction network was constructed to explore biological functions and identify potential therapeutic options. RESULTS: In the MR analysis, genetically predicted levels of complement C1q C chain (C1QC) were positively associated with the risk of autoimmune hepatitis (odds ratio 1.125, 95% confidence interval 1.018-1.244), while complement factor H-related protein 5 (CFHR5) was positively associated with the risk of primary sclerosing cholangitis (PSC;1.193, 1.048- 1.357). On the other hand, CFHR1 (0.621, 0.497-0.776) and CFHR2 (0.824, 0.703-0.965) were inversely associated with the risk of alcohol-related cirrhosis. There were also significant inverse associations between C8 gamma chain (C8G) and PSC (0.832, 0.707-0.979), as well as the risk of metabolic dysfunction-associated steatotic liver disease (1.167, 1.036-1.314). Additionally, C1S (0.111, 0.018-0.672), C7 (1.631, 1.190-2.236), and CFHR2 (1.279, 1.059-1.546) were significantly associated with the risk of hepatocellular carcinoma. Proteins from the complement regulatory networks and various liver diseaserelated proteins share common biological processes. Furthermore, potential therapeutic drugs for various liver diseases were identified through drug repurposing based on the complement regulatory network. CONCLUSION: Our study suggests that certain complement components, including C1S, C1QC, CFHR1, CFHR2, CFHR5, C7, and C8G, might play a role in non-viral liver diseases and could be potential targets for drug development.

Development and validation of machine-learning models for the difficulty of retroperitoneal laparoscopic adrenalectomy based on radiomics.

Objective: The aim is to construct machine learning (ML) prediction models for the difficulty of retroperitoneal laparoscopic adrenalectomy (RPLA) based on clinical and radiomic characteristics and to validate the models. Methods: Patients who had undergone RPLA at Shanxi Bethune Hospital between August 2014 and December 2020 were retrospectively gathered. They were then randomly split into a training set and a validation set, maintaining a ratio of 7:3. The model was constructed using the training set and validated using the validation set. Furthermore, a total of 117 patients were gathered between January and December 2021 to form a prospective set for validation. Radiomic features were extracted by drawing the region of interest using the 3D slicer image computing platform and Python. Key features were selected through LASSO, and the radiomics score (Rad-score) was calculated. Various ML models were constructed by combining Rad-score with clinical characteristics. The optimal models were selected based on precision, recall, the area under the curve, F1 score, calibration curve, receiver operating characteristic curve, and decision curve analysis in the training, validation, and prospective sets. Shapley Additive exPlanations (SHAP) was used to demonstrate the impact of each variable in the respective models. Results: After comparing the performance of 7 ML models in the training, validation, and prospective sets, it was found that the RF model had a more stable predictive performance, while xGBoost can significantly benefit patients. According to SHAP, the variable importance of the two models is similar, and both can reflect that the Rad-score has the most significant impact. At the same time, clinical characteristics such as hemoglobin, age, body mass index, gender, and diabetes mellitus also influenced the difficulty. Conclusion: This study constructed ML models for predicting the difficulty of RPLA by combining clinical and radiomic characteristics. The models can help surgeons evaluate surgical difficulty, reduce risks, and improve patient benefits.

Development and validation of nomograms to predict survival of primary adrenal lymphoma: a population-based retrospective study.

While it is known that accurate evaluation of overall survival (OS) and disease-specific survival (DSS) for patients with primary adrenal lymphoma (PAL) can affect their prognosis, no stable and effective prediction model exists. This study aimed to develop prediction models to evaluate survival. This study enrolled 5448 patients with adrenal masses from the SEER Program. The influencing factors were selected using the least absolute shrinkage and selection operator regression model (LASSO) and Fine and Gray model (FGM). In addition, nomograms were constructed. Receiver operating characteristic curves and bootstrap self-sampling methods were used to verify the discrimination and consistency of the nomograms. The independent influencing factors for PAL survival were selected by LASSO and FGM, and three models were built: the OS, DSS, and FGS (DSS analysis by FGM) model. The areas under the curve and decision curve analyses indicated that the models were valid. This study developed survival prediction models to predict OS and DSS of patients with PAL. The FGS model was more accurate than the DSS model in the short term. Above all, these models should offer benefits to patients with PAL in terms of the treatment modality choice and survival evaluation.

Brushite-coated Mg-Nd-Zn-Zr alloy promotes the osteogenesis of vertebral laminae through IGF2/PI3K/AKT signaling pathway.

Biodegradable magnesium (Mg) alloys have been extensively investigated in orthopedic implants due to their suitable mechanical strength and high biocompatibility. However, no studies have reported whether Mg alloys can be used to repair lamina defects, and the biological mechanisms regulating osteogenesis are not fully understood. The present study developed a lamina reconstruction device using our patented biodegradable Mg-Nd-Zn-Zr alloy (JDBM), and brushite (CaHPO4·2H2O, Dicalcium phosphate dihydrate, DCPD) coating was developed on the implant. Through in vitro and in vivo experiments, we evaluated the degradation behavior and biocompatibility of DCPD-JDBM. In addition, we explored the potential molecular mechanisms by which it regulates osteogenesis. In vitro, ion release and cytotoxicity tests revealed that DCPD-JDBM had better corrosion resistance and biocompatibility. We found that DCPD-JDBM extracts could promote MC3T3-E1 osteogenic differentiation via the IGF2/PI3K/AKT pathway. The lamina reconstruction device was implanted on a rat lumbar lamina defect model. Radiographic and histological analysis showed that DCPD-JDBM accelerated the repair of rat lamina defects and exhibited lower degradation rate compared to uncoated JDBM. Immunohistochemical and qRT-PCR results showed that DCPD-JDBM promoted osteogenesis in rat laminae via IGF2/PI3K/AKT pathway. This study shows that DCPD-JDBM is a promising biodegradable Mg-based material with great potential for clinical applications.

Accurate transcriptome-wide identification and quantification of alternative polyadenylation from RNA-seq data with APAIQ.

Alternative polyadenylation (APA) enables a gene to generate multiple transcripts with different 3' ends, which is dynamic across different cell types or conditions. Many computational methods have been developed to characterize sample-specific APA using the corresponding RNA-seq data, but suffered from high error rate on both polyadenylation site (PAS) identification and quantification of PAS usage (PAU), and bias toward 3' untranslated regions. Here we developed a tool for APA identification and quantification (APAIQ) from RNA-seq data, which can accurately identify PAS and quantify PAU in a transcriptome-wide manner. Using 3' end-seq data as the benchmark, we showed that APAIQ outperforms current methods on PAS identification and PAU quantification, including DaPars2, Aptardi, mountainClimber, SANPolyA, and QAPA. Finally, applying APAIQ on 421 RNA-seq samples from liver cancer patients, we identified >540 tumor-associated APA events and experimentally validated two intronic polyadenylation candidates, demonstrating its capacity to unveil cancer-related APA with a large-scale RNA-seq data set.

A nomogram for evaluation and analysis of difficulty in retroperitoneal laparoscopic adrenalectomy: A single-center study with prospective validation using LASSO-logistic regression.

Background: While it is known that inaccurate evaluation for retroperitoneal laparoscopic adrenalectomy (RPLA) can affect the surgical results of patients, no stable and effective prediction model for the procedure exists. In this study, we aimed to develop a computed tomography (CT) -based radiological-clinical prediction model for evaluating the surgical difficulty of RPLA. Method: Data from 398 patients with adrenal tumors treated by RPLA in a single center from August 2014 to December 2020 were retrospectively analyzed and divided into sets. The influencing factors were selected by least absolute shrinkage and selection operator regression model (LASSO). Additionally, the nomogram was constructed. A receiver operating characteristic curve was used to analyze the prediction efficiency of the nomogram. The C-index and bootstrap self-sampling methods were used to verify the discrimination and consistency of the nomogram. Result: The following 11 independent influencing factors were selected by LASSO: body mass index, diabetes mellitus, scoliosis, hyperlipidemia, history of operation, tumor diameter, distance from adrenal tumor to upper pole of kidney, retro renal fat area, hyperaldosteronism, pheochromocytoma and paraganglioma, and myelolipoma. The area under the curve (AUC) of the training set was 0.787, and 0.844 in the internal validation set. Decision curve analyses indicated the model to be useful. An additional 117 patients were recruited for prospective validation, and AUC was 0.848. Conclusion: This study developed a radiological-clinical prediction model proposed for predicting the difficulty of RPLA procedures. This model was suitable, accessible, and helpful for individualized surgical preparation and reduced operational risk. Thus, this model could contribute to more patients' benefit in circumventing surgical difficulties because of accurate predictive abilities.

The relationship between bladder storage function and frequent micturition after TURP.

PURPOSE: To investigate the relationship between preoperative bladder function and frequent micturition after transurethral resection of prostate in patients with benign prostatic hyperplasia. METHODS: We retrospectively included 80 eligible patients aged 54-87 years (mean age 69.8 years) who underwent transurethral resection of the prostate at our hospital from January 2019 to October 2021. Patients were divided into detrusor overactivity positive and negative groups, and according to bladder compliance, they were divided into: low (G1), normal (G2), and high (G3) bladder compliance groups. RESULTS: The incidence and score of postoperative frequent micturition in the detrusor overactivity positive group were higher than those in the detrusor overactivity negative group. The incidence and score of postoperative frequent micturition in the low bladder compliance group were higher than those in the normal and high bladder compliance groups. There was no significant difference in the score of frequent micturition between the normal and high bladder compliance groups. Multivariate logistic regression analysis indicated that frequent micturition was significantly correlated with detrusor overactivity, bladder compliance, maximum cystometric capacity, and maximum flow rate. CONCLUSION: This study confirmed that patients with abnormal bladder storage functions (detrusor instability and low bladder compliance) before transurethral resection of the prostate were likely to have frequent and severe urination after transurethral resection of the prostate. Therefore, preoperative urodynamic examination to evaluate the urinary storage function of patients with benign prostatic hyperplasia is of great significance to predict the occurrence and degree of postoperative frequent micturition.

Thrap3 promotes osteogenesis by inhibiting the degradation of Runx2.

The dysfunction of osteogenesis is a key character in the pathogenesis of osteoporosis, but the network of signaling mechanisms in controlling the differentiation of osteoblast remain unclear. Thrap3 has been proved participating in various biological process, especially in the differentiation of stem cells. Here, we demonstrate that Thrap3 could promote osteogenesis through the inhibition of the degradation of Runx2, which is a key molecular structure in early osteoblast differentiation. Furthermore, we found that the osteogenesis enhancing capacity of Thrap3 was caused by physically binding with Sox9, inhibiting the transcriptional activity of Sox9, and then decreasing the decomposition-promoted effect of Sox9 on Runx2. Our data shows that Thrap3 promotes osteoblast differentiation through the Thrap3-Sox9-Runx2 axis. What we found may help for further clarifying the molecular mechanism of osteogenic differentiation and give a new potential therapeutic target for osteoporosis.

Effects of 2,2',4'-trihydroxychalcone on the proliferation, metastasis and apoptosis of A549 human lung cancer cells.

The aim of the present study was to evaluate the antitumor effects of 2,2',4'-trihydroxychalcone (7a) on the A549 human lung cancer cell line. A549 cells were treated with different concentrations of 7a for different time periods. Cells without 7a were used as the negative control group. Cell proliferation, invasion, vasculogenic mimicry (VM) formation, heterogeneous adhesion and apoptosis were measured using Cell Counting Kit-8, Transwell invasion, VM, adhesion and flow cytometric assays, respectively. In addition, the expression of related proteins was determined using western blot analysis or ELISA. The present study found that 7a had a significant inhibitory effect on the survival rate of the A549 lung cancer cells but almost no effect on BEAS-2B human lung epithelial cells or human venous endothelial cells. The migration rate, VM length, invasion rate and heterogeneous adhesion number of cells treated with 7a significantly decreased as the concentration increased, while the apoptosis rate increased. Western blot analysis showed that 7a treatment significantly increased the expression levels of E-cadherin, cleaved poly (ADP-ribose) polymerase, Bax and caspase-3 and simultaneously decreased the expression levels of metalloproteinase-2/9, Bcl-2, phosphorylated (p)-PI3K, p-AKT, p-mTOR, vascular endothelial growth factor (VEGF), E-selectin and N-cadherin. At the same time, the ELISA results showed that the level of the pro-angiogenic factor VEGF in the culture media was reduced in the presence of 7a. In addition, 7a could also reduce the nuclear NF-κB protein expression, which could inhibit the gene transcription of tumor apoptosis and metastasis-related proteins. Therefore, 7a may exert inhibitory effects on A549 cells by inhibiting cell proliferation, migration, VM formation and heterogeneous adhesion, as well as by inducing apoptosis through the suppression of the PI3K/AKT/NF-κB signaling pathway; these findings suggested that 7a may be a promising agent for the treatment of lung cancer.

Perioperative factors influencing the difficulty of retroperitoneal laparoscopic adrenalectomy: a single-center retrospective study.

PURPOSE: Identifying patients in whom adrenalectomy may be more difficult can help with surgical decision-making. This study investigated the perioperative factors affecting the difficulty of retroperitoneal laparoscopic adrenalectomy (RLA). METHODS: Sixty-eight patients who underwent RLA at our hospital between December 1, 2020 and May 1, 2021 were included. The difficulty of RLA was assessed by operating time and intraoperative blood loss. We analyzed the relationship between surgical difficulty and patient sex, age, and body mass index, pathological type, tumor side, tumor size, distance from the lower pole of the adrenal tumor to the upper pole of the kidney (DAK), and distance from the lower pole of the adrenal tumor to the renal pedicle (DARP). RESULTS: Mean operating time was 105.38 ± 33.31 min and mean intraoperative blood loss was 32.28 ± 22.88 ml. Univariate linear regression analysis showed that age (P = 0.047), tumor size (P = 0.002), DAK (P = 0.002), and DARP (P < 0.001) were significantly correlated with a longer operating time. Univariate logistic regression analysis showed that DARP (P = 0.001), DAK (P = 0.001), tumor size (P = 0.002), and age (P = 0.033) were significantly correlated with a longer operating time. Multivariate logistic regression indicated that DARP (OR 5.341; 95% CI 1.704-16.739; P = 0.004), and tumor size (OR 4.433; 95% CI 1.434-13.709; P = 0.010) were independent predictors of operating time. CONCLUSION: Age, tumor size, DAK, and DARP were predictors of the difficulty of RLA. Older age, lower DARP and DAK, and a larger tumor size were associated with a longer operating time. DARP and tumor size were independent predictors of surgical difficulty.

Identification of carbohydrate peripheral epitopes important for recognition by positive-ion MALDI multistage mass spectrometry.

Carbohydrate sequences are important for various biological processes. It has recently been estimated to have 100,000-500,000 carbohydrate structures in mammalian glycome. However, the peripheral carbohydrate determinants on N- and O-glycoproteins, glycolipids, polysaccharides and secreted free sugars are limited in numbers. Among these blood-group-related antigens the ABO(H)- and Lewis-types are particularly important. Negative-ion MS/MS has been successfully used in assignment of these epitopes on free reducing sugars but cannot be applied to reduced sugars, e.g. O-glycans typically released from mucins as alditols, or in positive-ion detection of either reducing or reduced oligosaccharides. In the present study, we investigate the fragmentation features of permethylated reducing and reduced sugars under positive-ion conditions of multi-stage MALDI-MS, and propose the concept of epitope ion and epitope spectrum for determination of peripheral blood-group related epitopes on secreted human milk oligosaccharides and N-glycans as reducing sugars and O-glycans as reduced alditols in conjunction with MALDI-MS glycan profiling.

Polyvinyl alcohol degradation by Bacillus cereus RA23 from oil sludge sample.

A novel polyvinyl alcohol (PVA)-degrading strain Bacillus cereus RA23 was isolated from an oil sludge sample and environmental factors affecting its PVA degradation efficiency were optimized in detail. Inorganic nitrogen source, ammonium chloride (NH4Cl), was found to be the best nitrogen source and enhanced the PVA degradation rate greatly. The optimal medium for PVA biodegradation consisted of (g/L) PVA 1, NH4Cl 1, K2HPO4 1.6, MgSO4·7H2O 0.05, FeSO4·6H2O 0.02, CaCl2 0.05, NaCl 0.02. The optimal temperature and pH for PVA biodegradation by strain RA23 was 28 °C and 7.0, respectively, and 85% of 0.1% PVA was degraded after 5 days under these conditions. FTIR studies showed that the carboxylic acids (possibly including aldehyde or ketone) could be the intermediate product of PVA biodegradation. The investigation of strain RA23 for PVA degradation will provide important information to facilitate the removal of wastewater pollution in industrial zones.

Cryptochrome 1 Regulates Osteoblast Differentiation via the AKT Kinase and Extracellular Signal-Regulated Kinase Signaling Pathways.

The many circadian clock genes build up a network structure that controls physiological processes, such as the sleep cycle, metabolism, and hormone secretion. Cryptochrome 1 (CRY1), as one of the critical circadian proteins, is closely related to bone formation. However, the regulatory function of CRY1 in osteogenic differentiation remains unclear. In this study, we investigated the role of CRY1 in regulating proliferation and osteoblast differentiation in C3H10 and C2C12 cells after silencing Cry1 using short hairpin RNA interference. In vitro experiments confirmed that the expression level of CRY1 gradually increased during the osteogenic differentiation process, and Cry1 knockdown inhibited the proliferation and differentiation of osteoblastic cells. In addition, Cry1 knockdown inhibited the phosphorylation of AKT kinase (AKT) and extracellular signal-regulated kinase (ERK), which suppressed the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)-AKT and mitogen-activated protein kinase (MAPK)-ERK signaling pathways. Taken together, these findings show that CRY1 regulates the proliferation and differentiation of osteoblastic cells in an AKT and ERK-dependent manner.

Neolignan glycosides from Spiraea salicifolia and their inhibitory activity on pro-inflammatory cytokine interleukin-6 production in lipopolysaccharide-stimulated RAW 264.7 cells.

Phytochemical investigation of the ethanol extract from the roots of Spiraea salicifolia L. (Rosaceae) led to the isolation of one new neolignan glycoside, (7S,8R)-3,5-dimethoxy-4',7-epoxy-8,5'-neolignan-3',4,9,9'-tetraol 9-O-α-L-rhamnopyranoside (1), along with two known neolignan glycosides, (7S,8R)-3-methoxy-4',7-epoxy-8,5'-neolignan-3',4,9,9'-tetraol 9-O-α-L-rhamnopyranoside (2) and (7R,8S)-dihydrodehydrodiconiferyl alcohol 9'-O-ß-d-glucopyranoside (3). The structural characterisations of the isolated compounds were elucidated by spectroscopic data and comparison with the literatures reported on the chemical constituents from Juniperus communis var. depressa. All compounds were isolated for the first time from the Spiraea genus. Furthermore, the isolated compounds were subjected to assay inhibitory activity on pro-inflammatory cytokine interleukin-6 production in lipopolysaccharide-stimulated RAW 264.7 cells and all the compounds showed anti-inflammatory effects.

Cry 1 Regulates the Clock Gene Network and Promotes Proliferation and Migration Via the Akt/P53/P21 Pathway in Human Osteosarcoma Cells.

The many circadian clock genes buildup a network structure that controls physiological processes such as sleep cycle, metabolism and hormone secretion. A close relationship exists between circadian rhythm and cancers because cell cycle is affected by clock controlled genes (CCGs), including Cyclin D1, Cyclin A, Cyclin E and P21. The abnormal expression of the core circadian clock gene Cryptochrome 1 (Cry1) was found in many types of cancers. However, it is still unclear the exact mechanism of Cry1 dysregulation influences carcinogenesis and progression of cancers. In this study, we investigated the role of Cry1 in regulating proliferation and migration of Hos and U2os human osteosarcoma cells by silencing Cry1 using short hairpin RNA interference. Our data from in vitro and in vivo experiments confirmed that Cry1 knockdown enhanced proliferation and migration of osteosarcoma cells. Then, Cry2, Per1, Per2, Per3, Bmal1 and Clock were found up regulated, while Dec1, Dec2, CK1ε and Npas2 were downregulated at mRNA level. Besides, Akt/P53/P21 signaling was activated after Cry1 silencing and Akt was negatively phosphorylated along with Cry1 expression, while enhanced progression of osteosarcoma cells by Cry1 knockdown was reversed when Akt inhibitor treated. Furthermore, the rescue experiment verified the Akt/P53/P21 was downstream genes of Cry1 to control osteosarcoma progression. Taken together, these findings provide a new insight into how Cry1 regulates clock gene network and promotes proliferation and migration in a Akt dependent manner in human osteosarcoma cells.