Hepatitis B Virus Increases SphK1-S1P Synthesis by Promoting the Availability of the Transcription Factor USF1.

Hepatitis B virus (HBV) is the most common chronic viral infection globally, affecting ∼360 million people and causing about 1 million deaths annually due to end-stage liver disease or hepatocellular carcinoma. Current antiviral treatments rarely achieve a functional cure for chronic hepatitis B, highlighting the need for improved monitoring and intervention strategies. This study explores the role of the sphingosine kinase 1 (SphK1)-sphingosine-1-phosphate (S1P) axis in HBV-related liver injury. We investigated the association between serum S1P concentration and HBV DNA levels in chronic hepatitis B patients, finding a significant positive correlation. Additionally, SphK1 was elevated in liver tissues of HBV-positive hepatocellular carcinoma patients, particularly in HBsAg-positive regions. HBV infection models in HepG2-sodium taurocholate cotransporting polypeptide cells confirmed that HBV enhances SphK1 expression and S1P production. Inhibition of HBV replication through antiviral agents and the CRISPR-Cas9 system reduced SphK1 and S1P levels. Further, we identified the transcription factor USF1 as a key regulator of SphK1 expression during HBV infection. USF1 binds to the SphK1 promoter, increasing its transcriptional activity, and is upregulated in response to HBV infection. In vivo studies in mice demonstrated that HBV exposure promotes the expression of USF1 and SphK1-S1P. These findings suggest that the SphK1-S1P axis, regulated by HBV-induced USF1, could serve as a potential biomarker and therapeutic target for HBV-related liver injury.

Bergapten promotes bone marrow stromal cell differentiation into osteoblasts in vitro and in vivo.

Many recent studies have suggested that bergapten (BP), a class of native compound with numerous biological activities such as anti-resorptive properties, may exert protective effects against postmenopausal bone loss. However, it remains unknown whether BP regulates or improves the osteogenic function of bone marrow stromal cells (BMSCs) in the treatment and prevention of osteoporosis. In our study, BMSCs were cultured in osteogenic induction medium with the addition of BP for 2 weeks and an ovariectomized mouse model of osteoporosis was used to investigate the anti-resorptive effect of BP by gavage administration for 3 months. The concentrations of BP used were 0.1, 1, and 10 µmol/L in vitro and the gavage dose was 20 mg/kg/d. The result of our study indicated that BP promotes the expression of alkaline phosphatase (ALP) by BMSCs in vitro in a dose-dependent manner, as revealed by ALP staining. Runt-related transcription factor 2 and osteocalcin were up-regulated both in vitro and vivo, while osterix and collagen Iα1, assessed by immunofluorescence and immunohistochemistry, were correspondingly raised in the presence of BP in BMSCs in vitro. In addition, a protective effect of BP against ovariectomy-induced bone loss was found by distal femur micro-CT scanning, with improvements of bone metabolism parameters such as bone mineral density, trabecular number, and trabecular separation. Furthermore, WNT/ß-catenin signaling was activated in the presence of BP in BMSCs in osteogenic culture. Finally, BP promoted differentiation of BMSCs into osteoblasts by up-regulation of the WNT/ß-catenin pathway.

[Analysis of Volatile Components of Polypodium hastatum by GC-MS].

OBJECTIVE: To further reveal the chemical constituents of Polypodium hastatum, volatile components from this plant were investigated. METHODS: The volatile components were extracted under reflux from the whole plant of Polypodium hastatum, and then analyzed qualitatively and quantitatively by GC-MS. RESULTS: 60 volatile components were detected and of all components detected, the structures and relative contents of 34 volatile compounds were elucidated. CONCLUSION: In the volatile components identified, most are fatty acid esters, especially methyl and ethyl esters, which compose the major volatile chemical constituents of Polypodium hastatum.

[Expression of TLR8 in human cervical cancer HeLa cells and the effect of TLR8 agonist on the cell proliferation and apoptosis].

OBJECTIVE: To observe the expression of Toll-like receptor 8 (TLR8) in human cervical cancer cell-line HeLa cells, and the effects of TLR8 agonist CL075 on the survival and proliferation of HeLa cells. METHODS: PCR and RT-PCR were used to detect the expression of TLR8 in 13 cancer cell lines, and the expression of COX-2, Bcl-2, VEGF mRNA in the HeLa cells stimulated by TLR8 agonist CL075 were also measured by RT-PCR. Immunofluorescence technique was used to determine the exact location of TLR8 in the cells. The percentage of viable cells was determined by trypan blue exclusion after the HeLa cells were stimulated with TLR8 agonist CL075 (0.1 µg/ml, 0.5 µg/ml, 1.0 µg/ml, 2.5 µg/ml), and cell cycle and apoptosis were analyzed by flow cytometry, and the proliferation was measured by MTT. RESULTS: Compared with the other cancer cell lines, the expression of TLR8 in HeLa cells was the highest (703.7 ± 20.6). After stimulation by CL075, the cells had a remarkable increase of the percentage of cells in G(2)/M + S phases. In the control group, the percentage of cells in G(2)/M +S phases was (39.02 ± 2.33)%, whereas after stimulated with 1.0 µg/ml CL075, the percentage of cells in G(2)/M + S phases reached the highest ratio (57.67 ± 1.73)%, and the percentage of cells in G(2)/M + S phases had a less decrease after 2.5 µg/ml CL075 stimulation and the percentage was (56.14 ± 3.73)%. After the CL075 treatment, there was no significant changes of apoptosis compared with that of the control cells (P > 0.05), but after DDP treatment the apoptosis had a significant change (P < 0.01). After stimulation by 1.0 µg/ml CL075 for 24 h, no significant difference (P > 0.05) was found by MTT test, but a significant difference was found at 48 h and 72 h (P < 0.01). An increased expression of COX-2, Bcl-2 and VEGF mRNA was observed in HeLa cells after stimulation by TLR8 agonist CL075 for 24 h and 48 h (P < 0.05). CONCLUSIONS: Expression of TLR8 is significantly increased in HeLa cells. The proportion of cells at different phases has a significant change after CL075 stimulation, which may up-regulate the proliferation of HeLa cells. These data suggested that TLR8 agonist may influence the tumor development and TLR8 may become a potential target in the treatment for cervical cancer.

[Magnetic resonance imaging versus DR for whole spine imaging in patients with degenerative spinal disease].

OBJECTIVE: To assess the quality of whole spine images obtained by DR and magnetic resonance imaging (MRI) and analyze the whole spinal imaging sagittal parameters for standing DR and supine MRI. METHODS: Sixty-one patients aged 49.9∓17.6 years with degenerative spinal disease underwent both standing DR and supine MRI of the whole spine from November, 2010 to March, 2016. The image quality was retrospectively reviewed, and the cervical lordosis (CL), thoracic kyphosis (TK), lumbar lordosis (LL), sacral slope (SS), and sagittal vertical axis (SVA) were measured on the whole spinal lateral DR and middle sagittal MR images. RESULTS: Both the DR and MR whole spine images had a high quality (100%). The CL, TK, LL, SS, and SVA measured were 28.37mnplus;10.91 °, 29.98mnplus;8.96 °, 45.61mnplus;12.46 °, 34.38mnplus;9.05 °, and 17.20mnplus;26.39 mm on DR images and were 24.34mnplus;9.01 °, 21.22mnplus;8.13 °, 41.45mnplus;12.17 °, 37.45mnplus;8.19 °, and 36.51mnplus;12.44mm on MR images, respectively, showing significant differences in the measurements between the two modalities (P=0.000, 0.000, 0.000, 0.001, and 0.007, respectively). The correlation coefficient between DR and MR images for CL, TK, LL, SS, and SVA were 0.69, 0.68, 0.72, 0.51, and 0.27 (P=0.000, 0.000, 0.000, 0.000, and 0.034, respectively). CONCLUSION: Both standing DR and supine MR whole spine imaging can provide high-quality images. The CL, TK, LL, SS, and SVA measured on supine MR whole spine images are correlated with those on standing DR images but differ obviously. Supine MR imaging can not substitute standing DR examinations, and comprehensive assessment of degenerative spinal disease needs the combination of the two imaging techniques.

[Correlation between vertebral bone marrow fat and abdomen fat: a prospective study].

OBJECTIVE: To investigate the correlation between the lumbar bone marrow fat and abdominal fat. METHODS: A total of 68 individuals (32 men and 36 women, aged 21-74 years with a median of 49.5 years) were included in this study. All the subjects underwent spectroscopic examination of the third lumber vertebra with the single voxel method on a 1.5T MR scanner to measure the fat fraction (FF%). Quantitative CT was also performed for measurement of the abdomen subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT). The measurements were compared between subjects aged ≥50 years and those below 50 years, respectively,in male or female subjects. RESULTS: In male subjects, BMI, FF%, VAT or SAT showed no significant differences between the two age groups (P>0.05), and FF% was not correlated with BMI, VAT or SAT (r=0.109, 0.034, 0.066, respectively; P>0.05). In the female subjects, BMI, FF%, VAT and SAT differed significantly between the two age groups (P<0.05), and in ≥50 years group, FF% showed a positive correlation with VAT (r=0.499, P<0.05) but was not correlated with SAT (r=0.221, P>0.05); in<50 years group, FF% was not correlated with VAT or SAT (r=0.076, -0.067, respectively; P>0.05). CONCLUSION: FF% is positively correlated with VAT in female subjects aged beyond 50 years, but is not correlated with VAT or SAT in male subjects or in younger female subjects.