E3 ubiquitin ligase RNF114 promotes vesicular stomatitis virus replication via inhibiting type I interferon production.

As a single-stranded RNA virus, vesicular stomatitis virus (VSV) causes influenza-like clinical symptoms in infected individuals. Type-I interferon signaling pathway plays a vital role in inhibiting VSV replication. It has been shown that RNF114 (RING finger protein 114) acts as an E3 ubiquitin ligase to regulate the type-I interferon signaling pathway. In contrast, the effects of RNF114 from Chinese sturgeon or sea perch remain controversial. In the present study, we reported the effect of human RNF114 on VSV infection. Overexpression of RNF114 promoted VSV replication, while depletion of RNF114 reduced viral replication. We further found that RNF114 inhibited type-I interferon production via interacting with mitochondrial antiviral signaling protein (MAVS). Moreover, in vivo experiments demonstrated that RNF114 could also accelerate VSV replication and virus-induced inflammation in lung tissues. Collectively, our findings supported that RNF114 negatively regulated the type-I interferon signaling pathway during VSV replication, providing novel and favorable insights into clinical treatment of VSV infection.

Fully automated radiomic screening pipeline for osteoporosis and abnormal bone density with a deep learning-based segmentation using a short lumbar mDixon sequence.

BACKGROUND: Although lumbar bone marrow fat fraction (BMFF) has been demonstrated to be predictive of osteoporosis, its utility is limited by the requirement of manual segmentation. Additionally, quantitative features beyond simple BMFF average remain to be explored. In this study, we developed a fully automated radiomic pipeline using deep learning-based segmentation to detect osteoporosis and abnormal bone density (ABD) using a <20 s modified Dixon (mDixon) sequence. METHODS: In total, 222 subjects underwent quantitative computed tomography (QCT) and lower back magnetic resonance imaging (MRI). Bone mineral density (BMD) were extracted from L1-L3 using QCT as the reference standard; 206 subjects (48.8±14.9 years old, 140 females) were included in the final analysis, and were divided temporally into the training/validation set (142/64 subjects). A deep-learning network was developed to perform automated segmentation. Radiomic models were built using the same training set to predict ABD and osteoporosis using the mDixon maps. The performance was evaluated using the temporal validation set comprised of 64 subjects, along with the automated segmentation. Additional 25 subjects (56.1±8.8 years, 14 females) from another site and a different scanner vendor was included as independent validation to evaluate the performance of the pipeline. RESULTS: The automated segmentation achieved an outstanding mean dice coefficient of 0.912±0.062 compared to manual in the temporal validation. Task-based evaluation was performed in the temporal validation set, for predicting ABD and osteoporosis, the area under the curve, sensitivity, specificity, and accuracy were 0.925/0.899, 0.923/0.667, 0.789/0.873, 0.844/0.844, respectively. These values were comparable to that of manual segmentation. External validation (cross-vendor) was also performed; the area under the curve, sensitivity, specificity, and accuracy were 0.688/0.913, 0.786/0.857, 0.545/0.944, 0.680/0.920 for ABD and osteoporosis prediction, respectively. CONCLUSIONS: Our work is the first attempt using radiomics to predict osteoporosis with BMFF map, and the deep-learning based segmentation will further facilitate the clinical utility of the pipeline as a screening tool for early detection of ABD.

Kruppel-like factor 10 protects against acute viral myocarditis by negatively regulating cardiac MCP-1 expression.

Viral myocarditis (VMC) is a cardiac disease associated with myocardial inflammation and injury induced by virus infection. Cardiomyocytes have recently been regarded as key players in eliciting and modulating inflammation within the myocardium. Kruppel-like factor 10 (KLF10) is a crucial regulator of various pathological processes and plays different roles in a variety of diseases. However, its role in VMC induced by coxsackievirus B3 (CVB3) infection remains unknown. In this study, we report that cardiac KLF10 confers enhanced protection against viral myocarditis. We found that KLF10 expression was downregulated upon CVB3 infection. KLF10 deficiency enhanced cardiac viral replication and aggravated VMC progress. Bone marrow chimera experiments indicated that KLF10 expression in nonhematopoietic cells was involved in the pathogenesis of VMC. We further identified MCP-1 as a novel target of KLF10 in cardiomyocytes, and KLF10 cooperated with histone deacetylase 1 (HDAC1) to negatively regulate MCP-1 expression by binding its promoter, leading to activation of MCP-1 transcription and recruitment of Ly6Chigh monocytes/macrophages into the myocardium. This novel mechanism of MCP-1 regulation by KLF10 might provide new insights into the pathogenesis of VMC and a potential therapeutic target for VMC.

Vitamin D Provides Benefit Based on the Proinflammatory Effects of Homocysteine in Elderly Patients With Type 2 Diabetes Mellitus.

PURPOSE: It is unclear whether vitamin D provides any benefit against the pro-inflammatory effects of homocysteine in elderly patients with type 2 diabetes mellitus (T2DM). METHODS: We compared lymphocyte counts for CD3, CD19, CD4, and CD8 subsets between elderly (age ≥65 years) T2DM patients (n = 5098) and nondiabetes control subjects (n = 20,590) based on the serum concentrations of homocysteine and total vitamin D (calcidiol + calcifediol [total vitamin D, TVD]; <20, 20-30, and >30 ng/mL). FINDINGS: Significant variation in CD19 (P = 0.019), CD4 (P = 0.015), and CD8 (P < 0.001) were associated with serum TVD in T2DM patients with homocysteine ≤15 µmol/L, whereas CD3 (P = 0.003) and CD8 (P = 0.019) varied in control subjects with homocysteine ≤15 µmol/L. In T2DM patients with high homocysteine (>15 µmol/L) levels, significant variation based on serum TVD occurred in CD19 only (P = 0.024), whereas CD3 (P = 0.016) and CD4 (P = 0.001) varied in control subjects with high homocysteine concentrations. IMPLICATIONS: Serum TVD influences variation in CD3, CD19, CD4, and CD8 lymphocyte subsets based on the serum homocysteine concentration in elderly T2DM patients and nondiabetic individuals with moderate to high homocysteine concentrations. The effect of TVD is partially attenuated in individuals with high homocysteine concentrations, with greater attenuation occurring in patients with T2DM. Differences in the variation of lymphocyte subsets between nondiabetes subjects with moderate homocysteine concentrations and those with high homocysteine concentrations constitute a shift from CD8-positive cells to CD4-positive cells, suggesting a change in TH1/TH2 balance based on TVD and homocysteine concentrations that is absent in diabetes cases with high homocysteine concentrations.

Lithium chloride confers protection against viral myocarditis via suppression of coxsackievirus B3 virus replication.

Viral myocarditis (VMC) is a type of inflammation affecting myocardial cells caused by viral infection and has been an important cause of dilated cardiomyopathy (DCM) worldwide. Type B3 coxsackievirus (CVB3), a non-enveloped positive-strand RNA virus of the Enterovirus genus, is one of most common agent of viral myocarditis. Till now, effective treatments for VMC are lacking due to lack of drugs or vaccine. Lithium chloride (LiCl) is applied in the clinical management of manic depressive disorders. Accumulating evidence have demonstrated that LiCl, also as an effective antiviral drug, exhibited antiviral effects for specific viruses. However, there are few reports of evaluating LiCl's antiviral effect in mice model. Here, we investigated the inhibitory influence of LiCl on the CVB3 replication in vitro and in vivo and the development of CVB3-induced VMC. We found that LiCl significantly suppressed CVB3 replication in HeLa via inhibiting virus-induced cell apoptosis. Moreover, LiCl treatment in vivo obviously inhibited virus replication within the myocardium and alleviated CVB3-induced acute myocarditis. Collectively, our data demonstrated that LiCl inhibited CVB3 replication and negatively regulated virus-triggered inflammatory responses. Our finding further expands the antiviral targets of LiCl and provides an alternative agent for viral myocarditis.

MCPIP1 inhibits Hepatitis B virus replication by destabilizing viral RNA and negatively regulates the virus-induced innate inflammatory responses.

Monocyte chemotactic protein-induced protein 1 (MCPIP1) is an inflammatory regulator in immune response. Recently, MCPIP1 has also been identified as a host antiviral factor against certain virus infection including human immunodeficiency virus, dengue virus and hepatitis C virus. However, whether MCPIP1 could restrict the replication of hepatitis B virus (HBV), a DNA pararetrovirus belonging to Hepadnaviridae family, has not been investigated. In this study, we found that MCPIP1 expression was up-regulated in mouse livers upon acute HBV replication and in HBV-replicated hepatoma cells or HBV-stimulated macrophages. Enforced MCPIP1 expression by hydrodynamic DNA injection in vivo significantly inhibited HBV replication in the mouse livers. Then in vitro studies by overexpression or knockdown assays in cell-lines identified the direct antiviral effect of MCPIP1 on HBV replication. RNA immunoprecipitation and decay assay further suggested that MCPIP1 potently restricted HBV replication through directly binding viral RNA and degrading RNA via its RNase activity, but not deubiquitinase activity. Moreover, we further verified that MCPIP1 negatively regulated HBV-induced proinflammatory cytokines, such as IL-1ß, TNF-α and IL-6 in macrophages. Taken together, our data expand MCPIP1's range of viral targets to DNA virus and also demonstrate the negative regulatory role of MCPIP1 in suppressing virus-induced inflammatory response, suggesting MCPIP1 as a potential therapeutic target for treating HBV-related diseases via inducing a host defense against HBV and reducing inflammatory injury meanwhile.

Fatty infiltration of paraspinal muscles is associated with bone mineral density of the lumbar spine.

A total of 88 subjects were enrolled to investigate the relationship between paraspinal muscle fatty infiltration and lumbar bone mineral density (BMD) using chemical shift encoding-based water-fat MRI and quantitative computed tomography (QCT), respectively. A moderate inverse correlation between paraspinal muscle proton density fat fraction and lumbar QCT-BMD was found with age, sex, and BMI controlled. PURPOSE: To investigate the relationship between paraspinal muscle fatty infiltration and lumbar bone mineral density (BMD). METHODS: A total of 88 subjects were enrolled in this study (52 females, 36 males; age, 46.6 ± 14.2 years old; BMI, 23.2 ± 3.49 kg/m2). Proton density fat fractions (PDFF) of paraspinal muscles (erector spinae, multifidus, and psoas) were measured at L2/3, L3/4, and L4/5 levels using chemical shift encoding-based water-fat MRI. Quantitative computed tomography (QCT) was used to assess BMD of L1, L2, and L3. The differences in paraspinal muscle PDFF among subjects with normal bone density, osteopenia, and osteoporosis were tested using one-way ANOVA. The relationship between paraspinal muscle PDFF and QCT-BMD was analyzed using linear regression with age, sex, and BMI variables. RESULTS: PDFF of the erector spinae, multifidus, and psoas of subjects with normal bone density were all significantly less than those with osteopenia and those with osteoporosis (all p < 0.001). There was an inverse correlation between paraspinal muscle PDFF and BMD after controlling for age, sex, and BMI (standardized beta coefficient, - 0.21~- 0.29; all p < 0.05). CONCLUSIONS: Paraspinal muscle fatty infiltration increased while lumbar BMD decreased after adjusting for age, sex, and BMI. Paraspinal muscles and vertebrae are interacting tissues. Paraspinal muscle fatty infiltration may be a marker of low lumbar BMD. Chemical shift imaging is an efficient and fast quantitative method and can be easily added to the clinical protocol to measure paraspinal muscle PDFF when the patient underwent the routine lumbar MRI with low-back pain.

Prediction of Abnormal Bone Density and Osteoporosis From Lumbar Spine MR Using Modified Dixon Quant in 257 Subjects With Quantitative Computed Tomography as Reference.

BACKGROUND: Bone marrow fat increases when bone mass decreases, which could be attributed to the fact that adipogenesis competes with osteogenesis. Bone marrow fat has the potential to predict abnormal bone density and osteoporosis. PURPOSE: To investigate the predictive value of using vertebral bone marrow fat fraction(BMFF) obtained from modified Dixon(mDixon) Quant in the determination of abnormal bone density and osteoporosis. STUDY TYPE: Prospective. POPULATION: 257 subjects (age: 20-79 years old; BMI: 16.6-32.9 kg/m2 ;181 females,76 males) without known spinal tumor, history of trauma, dysplasia, spinal surgery or hormone therapy. FIELD STRENGTH/SEQUENCE: 3.0T/mDixon. ASSESSMENT: BMFF was measured at the L1, L2 and L3 vertebral body on fat fraction maps of the lumbar spine. Bone mineral density (BMD) was obtained using quantitative computed tomography, which served as the reference standard. STATISTICAL TESTS: The BMFF between the three groups (normal bone density, osteopenia and osteoporosis) was tested using one-way analysis of variance in SPSS. The correlation and partial correlation of BMFF and BMD were analyzed before and after controlling for age, sex and BMI. Logistic regression analysis using independent training and validation data was conducted to evaluate the performance of predicting abnormal BMD or osteoporosis using BMFF. RESULTS: There was a significant difference in vertebral BMFF between the three groups (P < 0.001). Moderate inverse correlation was found between vertebral BMFF and BMD after controlling age, sex and BMI (r = -0.529; P < 0.001). The mean area under the curve, sensitivity, specificity and negative predictive value (NPV) for predicting abnormal bone density were 0.940, 0.877, 0.896, and 0.890, respectively. The corresponding results for predicting subjects with osteoporosis were 0.896, 0.848, 0.853, and 0.969, respectively. DATA CONCLUSION: mDixon Quant is a fast, simple, noninvasive and nonionizing method to access vertebral BMFF and has a high predictive power for identifying abnormal bone density and osteoporosis. LEVEL OF EVIDENCE: 1 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2019;49:390-399.

[Changes in shape and signal intensity of high intensity zone in lumbar intervertebral discs on magnetic resonance images: a longitudinal study].

OBJECTIVE: To investigate changes over time in the shape and signal intensity of high intensity zone (HIZ) in the lumbar intervertebral discs on magnetic resonance images in patients with low back pain. METHODS: The imaging data were collected from 27 patients with low back pain, who underwent lumbar magnetic resonance (MR) imaging examinations that identified HIZ lesions and received follow-up MR examinations at least 1.5 years later over the period from January 2009 to January 2017. The initial and follow-up MR T2WI images of the patients were read by two experienced radiologists to categorize the changes in the shape of the HIZ lesions into enlarged, unchanged, shrunk, and disappeared. The volume and signal/cerebrospinal fluid signal intensity (T2/CSF) ratio of the HIZ were measured on sagittal MR images using ImageJ software. RESULTS: Of the 43 HIZ lesions found in the initial examinations, 22 (51.2%) remained unchanged in the follow-up examinations, 10 (21.3%) were enlarged, 9 (20.9%) shrank, and 2 (23.3%) disappeared. The follow-up examinations revealed 4 new HIZ lesions in the intervertebral discs. The volumes of these lesions did not vary significantly in the follow-up examinations (P=0.653), but the T2/CSF ratio was significantly higher in the follow-up than in the initial examinations (P=0.043). CONCLUSIONS: After observation for an average of 3 years and 3 months, most of the HIZ lesions in the lumbar intervertebral discs of the patients with low back pain remained stable in shape, but their signal intensity on MR images increased.

[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.

[Multiple linear regression and ROC curve analysis of the factors of lumbar spine bone mineral density].

OBJECTIVE: To investigate the correlation between the lumbar vertebra bone mineral density (BMD) and age, gender, height, weight, body mass index, waistline, hipline, bone marrow and abdomen fat, and to explore the key factor affecting the BMD. METHODS: A total of 72 cases were randomly recruited. All the subjects underwent a spectroscopic examination of the third lumber vertebra with single-voxel method in 1.5T MR. Lipid fractions (FF%) were measured. Quantitative CT were also performed to get the BMD of L3 and the corresponding abdomen subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT). The statistical analysis were performed by SPSS 19.0. RESULTS: Multiple linear regression showed except the age and FF% showed significant difference (P<0.05) , the gender, height, weight, BMI, waistline, hipline, SAT and VAT showed no significant difference (P>0.05). The correlation of age and FF% with BMD was statistically negatively significant (r=-0.830, -0.521, P<0.05). The ROC curve analysis showed that the sensitivety and specificity of predicting osteoporosis were 81.8% and 86.9%, with a threshold of 58.5 years old. And it showed that the sensitivety and specificity of predicting osteoporosis were 90.9% and 55.7%, with a threshold of 52.8% for FF%. CONCLUSION: The lumbar vertebra BMD was significantly and negatively correlated with age and bone marrow FF%, but it was not significantly correlated with gender, height, weight, BMI, waistline, hipline, SAT and VAT. And age was the critical factor.

[Correlation of vertebral body deformity and adjacent disc degeneration in patients with old thoracolumbar compression fractures].

OBJECTIVE: To assess the correlation between vertebral body deformity and degeneration of the adjacent intervertebral discs in patients with old thoracolumbar compression fractures. METHODS: Seventy-one patients who had been conservatively treated after single segment thoracolumbar compression fractures between April, 2011 and May, 2014 were enrolled in this study. Both radiographic and magnetic resonance (MR) images of the thoracolumbar segment were obtained. The involved vertebral body deformity was rated on radiography according to the Genant criterion, and the degeneration of the adjacent cephalic and caudal discs was assessed on MR images using the Oner and Pfirrmann classification schemes, respectively. The relationship between vertebral body deformity and adjacent disc changes was assessed using correlation analysis, and the changes in the adjacent cranial and caudal discs was compared. RESULTS: The Genant classification of the involved vertebral bodies was moderately correlated with Oner morphological scores (r=0.48, P<0.01), but not with the Pfirrmann signal scores of the adjacent cephalic discs or with the Genant or Pfirrmann scores of the adjacent caudal discs (P>0.05). The Oner classification of the adjacent cephalic discs was higher than that of the adjacent caudal discs (P<0.01), but their Pfirrmann classification did not differ significantly. CONCLUSION: The deformity of vertebral body affects the adjacent cephalic discs proportionally but not the adjacent caudal discs.

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.

[Three-dimensional isotropic magnetic resonance imaging of the ankle joint].

OBJECTIVE: To compare the image quality of isotropic 3-dimensional fast spin echo (3D-FSE), 3D fast field echo (3D-FFE), and 2D fast spin echo (2D-FSE) sequences in magnetic resonance imaging (MRI) of the anatomical structure of the ankle joint. METHODS: The ankle joints of 10 healthy volunteers were examined with isotropic 3D-FSE, 3D-FFE and 2D-FSE sequences using a 1.5T MR scanner and 3D reconstruction. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of the tissues were measured. Two radiologists evaluated the image quality of the 3 sequences using a 5-point Likert scale in a double-blinded manner. RESULTS: The 3D-FSE sequences resulted in the highest SNRs for all the tissues and the highest CNRs for differentiation between cartilage and marrow, between muscle and tendon, and between tendon and fluid. In the estimation of image quality for cartilages, 3D-FFE had the highest score followed by 3D-FSE, and the latter had the highest score among the 3 sequences in displaying the tendon. CONCLUSION: 3D-FSE sequence has a high performance in displaying the anatomical structures of complex joints especially for cartilage, ligament, and tendon tissues.

[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.

[Magnetic resonance imaging for pelvic bone marrow fat deposition in patients with ankylosing spondylitis].

OBJECTIVE: To evaluate the diagnostic value of pelvis bone marrow fat depositions (BMFD) displayed by magnetic resonance imaging (MRI) in patients with ankylosing spondylitis (AS). METHODS: Eighty-eight subjects undergoing pelvic MRI examinations were enrolled in this study, including 44 with clinically confirmed AS (39 male and 5 female patients with a mean age of 26.41∓8.09 years) and 44 control subjects without AS (37 male and 7 female subjects with a mean age of 29.32∓7.31 years). The incidence of BMFD in the bilateral sacroiliac (SI) joints and acetabulum were compared between the two groups. The distribution features of BMFD of the periarticular cancellous bone marrow in the pelvis and in other regions of the pelvis were analyzed for the AS patients, and the incidence of BMFD was determined in different stages of sacroiliitis and hip arthritis. RESULTS: The incidence of BMFD in the SI joints and acetabulum was significantly higher in the AS patients than in the control subjects (P<0.01); The incidence of BMFD was significantly higher in the periarticular cancellous bone marrow than in the other positions of pelvis (P<0.01). The incidence of BMFD ranged from 40.0% to 45.9% in early stages of sacroiliitis, significantly lower than the incidence in later stages (58.3%-73.1%, P<0.01); the incidence showed no difference between different stages of hip arthritis (P>0.01). CONCLUSIONS: AS patients have a higher incidence of BMFD in the pelvis than control subjects. BMFD is distributed mainly under the articular surface, seen throughout the stages of AS, indicating that BMFD is an important pathological change of the bone marrow in AS to potentially allow early diagnosis of AS.

[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.