Effects of ß-carotene supplementation in the diet of laying breeder hens on the growth performance and liver development of offspring chicks.

This experiment was conducted to evaluate the growth performance, growth regulating factors, and liver morphology of chicks hatched from egg-laying breeding hens dietary supplemented with additives (ß-carotene). Hy-line breeding hens were allocated into three groups with three replicates/group. The dietary treatments were as follows: basal diet as a control (Con), basal diet supplemented with 120 (ßc-L) or 240 (ßc-H) mg/kg of ß-carotene diet. After 6 weeks, the eggs were collected and incubated. The hatched chicks were fed the same diet. The results showed that chicks in the ßc-L group increased in body weight at 21 days (p < 0.01). At 42 days, chicks in the ßc-H group showed a significant increase in tibia length (p < 0.05). The liver index increased in the ßc-L and ßc-H groups at 7 days (p < 0.05). Serum HGF (7, 14, 21, and 42 days) and leptin (14 days) were significantly increased in the group supplemented with ßc. Hepatic GHR (14 days), IGF-1R (14 days), and LEPR (21 days) mRNA expression were significantly increased. In addition, there was an increase in PCNA-positive cells in the liver of chicks in the ßc group. In conclusion, the addition of ß-carotene to the diet of laying breeder hens was more advantageous in terms of growth performance and liver development of the offspring.

miR-96-5p is the tumor suppressor in osteosarcoma via targeting SYK.

BACKGROUND: Osteosarcoma is a multiple malignant tumor in adolescents. MicroRNAs (MiRNAs) have been found to express abnormally in OS tissues and are considered as potential targets for OS prognosis and treatment. METHODS: MiR-96-5p and SYK expression in clinical samples, osteoblast and OS cell lines were detected. The changes of cell proliferation, apoptosis, adhesion and metastasis of OS cells were detected by CCK-8, BrdU, caspase-3 activity and transwell assay. Dual luciferase report analysis and RNA pull-down were used to confirm binding relation of miR-96-5p and SYK. RESULTS: MiR-96-5p was increased in OS tissue and cells. Moreover, miR-96-5p inhibits proliferation, adhesion and migration of HOS and Saos-2 cells, and promotes cell apoptosis. SYK has been identified to be targeted by miR-96-5p. Overexpressed SYK inhibits the suppressive impact of miR-96-5 on OS cells. CONCLUSION: MiR-96-5p may function as an effective target in OS treatment.

Matching correction of main and compensatory curves is critical for immediate postoperative coronal balance in correction of severe adult idiopathic scoliosis.

PURPOSE: To analyze the correlation between immediate postoperative coronal imbalance and the matching degree of the correction rates of the main curve and compensatory curves in the surgical treatment of severe adult idiopathic scoliosis. METHODS: Patients were categorized into three types based on the preoperative coronal balance status (type A = balance, type B = shifted to cave side and type C = shifted to convex side), and each type was further divided into two subgroups based on the postoperative coronal balance status (balance and imbalance). Different coronal parameters before and after operations were calculated and compared. RESULTS: The rate of postoperative CIB was highest in type C patients (53.8%) and lowest in type A patients (31.5%). To avoid postoperative CIB, the value of the postoperative CRmain should fall within the range of 1.001 × CRcomp ± 2.84% in type A patients, 1.112 × CRcomp + 3.3% ± 5.02% in type B patients and 0.907 × CRcomp - 2.5% ± 4.38% in type C patients. CONCLUSION: Mismatch between the correction rates of the main curve and compensation curves is a critical cause of immediate postoperative CIB. The relatively equal correction of the main curve and compensatory curves is essential for type A patients to achieve postoperative coronal balance, while the correction rate of the main curve should be higher than the compensatory curves in type B patients and vice versa in type C patients. Three formulas for the three different types were developed to provide helpful guidance information for surgical planning.

Association between TNF-α polymorphisms and the risk of upper gastrointestinal bleeding induced by aspirin in patients with coronary heart disease.

OBJECTIVE: To investigate the correlation of tumor necrosis factor α (TNF-α) polymorphisms with upper gastrointestinal bleeding (UGIB) induced by enteric-coated aspirin in coronary heart disease (CHD) patients. METHODS: In total, 154 CHD patients taking enteric-coated aspirin were enrolled in this study. Patients were divided into the UGIB group (n = 57) and non-UGIB group (n = 97) based on the presence or absence of signs of UGIB, respectively. TNF-α polymorphism (-857C > T, -863C > A, and -1031T > C) genotyping was performed using polymerase chain reaction (PCR) amplification with sequence-specific primers (PCR-SSP). RESULTS: Patients who had the CC genotype and C allele of -1031T > C exhibited a significantly increase risk of UGIB after receiving enteric-coated aspirin (CC vs. TT: odds (OR) (95% confidence interval (CI)): 7.568 (1.527-37.49), P = 0.005; C vs. T: OR (95% CI): 1.852 (1.036-3.312), P = 0.036). Patients who had CA and CA + AA genotypes and the A allele of -863C > A also exhibited an increased risk of aspirin-induced UGIB (CA vs. CC: OR (95% CI): 2.415 (1.143-5.101), P = 0.019: CA + AA vs. CC: OR (95% CI): 2.218 (1.123-4.381), P = 0.021; A vs. C: OR (95% CI): 1.788 (1.039-3.078), P = 0.035). However, the TNF-α -857 C > T polymorphism was unrelated to the induction of UGIB by enteric-coated aspirin in CHD patients (P > 0.05). In addition, the haplotypes of CCC (-1031T > C, -863C > A, and -857C > T) markedly reduced the risk of aspirin-induced UGIB in CHD patients. CONCLUSION: TNF-α -863A and -1031C increased the risk of UGIB induction by enteric-coated aspirin in CHD patients, whereas TNF-α -857C > T was not correlated with the UGIB risk.

Blood concentration, efficacy, and adverse events of phenobarbital: A prospective study in rural China.

OBJECTIVE: This study evaluated the relationship between blood concentration of phenobarbital (PB) and its efficacy as well as adverse events in people with epilepsy in rural China. METHODS: People with epilepsy being treated with PB monotherapy were recruited and followed up for averagely 2.5 years. Data of clinical characteristics were collected using a standardized questionnaire by face-to-face interviews both at baseline and follow-up. Plasma concentration of PB was detected by the high-performance liquid chromatography. RESULTS: Data on treatment response and PB blood concentration was obtained from 225 subjects. Among them, 119 (52.9%) were recognized as effective cases and 106 (47.1%) as ineffective cases. In the effective group, the blood concentration of 95% subjects ranged from 1.22 µg/ml to 41.36 µg/ml with a median at 13.18 µg/ml (IQR = 8.32-20.19 µg/ml). The PB concentration of 95% of the subjects in the ineffective group ranged from 2.73 µg/ml to 70.16 µg/ml with a median at 19.80 µg/ml (IQR = 11.30-30.40 µg/ml), which was significantly higher than that of the effective group (p < 0.001). Multivariate logistic regression analysis showed that PB concentration ≥26.38 µg/ml was related to a 4.5-fold (95% confidence interval [CI], 1.85-11.08) higher risk of inefficacy. A receiver operation characteristic curve was performed to determine the cutoff value of concentration for PB efficacy at 19.02 µg/ml. SIGNIFICANCE: Blood concentration may be an important indicator for clinical decision making when PB monotherapy cannot achieve a good efficacy and more attention should be paid on it in clinical practice especially in resource-poor settings.

Incidence of sudden unexpected death in epilepsy in community-based cohort in China.

OBJECTIVE: Sudden unexpected death in epilepsy (SUDEP) is associated with the high premature mortality observed among people with epilepsy. It is, however, considered a rare event in China, probably because of lack of awareness and limitation of studies in the country. We aimed to provide some initial estimation of the burden of SUDEP in China. METHODS: We established a large Chinese community-based cohort of people with epilepsy between January 2010 and December 2011. For any participant who died during follow-up, detailed information on cause of death was obtained using a specifically designed Verbal Autopsy Questionnaire. All cases were reviewed by a multidisciplinary expert panel and reinvestigated if necessary. Sudden unexpected death in epilepsy incidence rates were estimated and case details provided. RESULTS: The cohort consisted of 1562 people and during a median 5years follow-up, 72 deaths were reported. The all-causes death incidence was 11.23 (95% CI 8.86-14.07) per 1000 person-years. Fifteen died suddenly and unexpectedly in a reasonable state of health in the week preceding death. We recorded detailed information of these 15 deaths. Thirteen were considered to be probable SUDEP and two possible SUDEP. The incidence of probable SUDEP was 2.03 (95% CI 1.13-3.38) per 1000 person-years, and the incidence of all suspected (probable and possible) SUDEP was 2.34 (95% CI 1.36-3.77) per 1000 person-years. SIGNIFICANCE: The incidence of SUDEP was relatively high among Chinese people with epilepsy when compared with that in previous community-based studies from high-income countries. The burden of SUDEP in China requires further assessments.

Nutritional supplementation of breeding hens may promote embryonic development through the growth hormone-insulin like growth factor axis.

The late stage of embryo development is a crucial period of metabolic changes, with rapid organ development requiring a substantial supply of nutrients. During this phase, maternal nutritional levels play a vital role in the growth, development, and metabolism of the offspring. In this study, we added 2 doses of ß-carotene (ßc) (120 mg/kg and 240 mg/kg) to the daily diet of Hailan Brown laying hens to investigate the impact of maternal nutritional enrichment on embryo development. Maternal nutrition supplementation significantly increased the expression of chicken embryo liver index, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and hepatocyte growth factor (HGF) in serum. At the same time, the expression of GH/growth hormone receptor (GHR), IGF-1 mRNA, and Proliferating Cell Nuclear Antigen (PCNA) protein in the liver was upregulated, indicating that maternal nutrition intervention may promote chicken embryo liver development through the GH-IGF-1 axis. Transcriptome sequencing results showed that differential genes in liver after maternal nutritional supplementation with ß-carotene were enriched in pathways related to cell proliferation and metabolism. Consequently, we postulated that maternal ß-carotene supplementation might operate via the GH-IGF-1 axis to regulate the expression of genes involved in growth and development, thereby promoting liver development. These results contribute to formulating more effective poultry feeding strategies to promote offspring growth and development.

Long-term outcome of phenobarbital treatment for epilepsy in rural China: a prospective cohort study.

PURPOSE: To evaluate the long-term outcome of phenobarbital treatment for convulsive epilepsy in rural China, and to explore factors associated with overall seizure outcomes. METHODS: We carried out follow-up assessments of people who took part in an epilepsy community management program conducted in rural counties of six provinces in China. People with convulsive epilepsy who were previously untreated (or on irregular treatment) were commenced on regular treatment with phenobarbital. Information was collected using a standardized questionnaire by face-to-face interviews of the individuals (and their families where necessary). Information collected included treatment status, medication change, seizure frequency, and mortality. KEY FINDINGS: Among the 2,455 people who participated in the original program, outcomes were successfully ascertained during the follow-up assessment in 1986. Among them, 206 had died. Information on treatment response was obtained in 1,780 (56% male; mean age 33.9 years, range 3-84; mean duration of follow-up 6.4 years). Among them, 939 (53%) were still taking phenobarbital. The most common reasons for stopping phenobarbital were seizure freedom or substantial seizure reduction, socioeconomic reasons, and personal preference. Four hundred fifty-three individuals (25%) became seizure-free for at least 1 year while taking phenobarbital, 88% of whom did so at daily doses of 120 mg or below. Four hundred six (23%) reported adverse events, which led to withdrawal of phenobarbital in <1%. The most common adverse effects were malaise/somnolence (7.4%), dizziness (3%), and lethargy (2.6%). At the follow-up assessment, 688 (39%) individuals had been seizure free for at least the previous year. People with persistent seizures had significantly longer duration of epilepsy and higher number of seizures in the 12 months before treatment. People who were taking AED treatment irregularly at recruitment were less likely to become seizure-free. SIGNIFICANCE: We observed long-term benefits of regular treatment with phenobarbital for convulsive epilepsy in rural China. One hundred years after the discovery of its antiepileptic effect, phenobarbital is still playing an important role in the management of epilepsy.

Premature mortality risk in people with convulsive epilepsy: long follow-up of a cohort in rural China.

PURPOSE: Detailed data on the mortality of epilepsy are still lacking from resource-poor settings. We conducted a long-term follow-up survey in a cohort of people with convulsive epilepsy in rural areas of China. In this longitudinal prospective study we investigated the causes of death and premature mortality risk among people with epilepsy. METHODS: We attempted to trace all 2,455 people who had previously participated in a pragmatic assessment of epilepsy management at the primary health level. Putative causes of death were recorded for those who died, according to the International Classification of Diseases. We estimated proportional mortality ratios (PMRs) for each cause, and standardized mortality ratios (SMRs) for each age-group and cause. Survival analysis was used to detect risk factors associated with increased mortality. KEY FINDINGS: During 6.1 years of follow-up there were 206 reported deaths among the 1,986 people with epilepsy who were located. The highest PMRs were for cerebrovascular disease (15%), drowning (14%), self-inflicted injury (13%), and status epilepticus (6%), with probable sudden unexpected death in epilepsy (SUDEP) in 1%. The risk of premature death was 2.9 times greater in people with epilepsy than in the general population. A much higher risk (SMRs 28-37) was found in young people. Duration of epilepsy and living in a waterside area were independent predictors for drowning. SIGNIFICANCE: Drowning and status epilepticus were important, possibly preventable, causes of death. Predictors of increasing mortality suggest interventions with efficient treatment and education to prevent premature mortality among people with epilepsy in resource-poor settings.

[Correlation of lumbar disc degeneration and spinal-pelvic sagittal balance].

OBJECTIVE: To elucidate the relationship between spino-pelvic sagittal balance parameters and lumbar intervertebral disc degeneration of each segment through retrospective analysis in lumbar degeneration patients. METHODS: Retrospective analysis was conducted for the follow-up data in 126 patients with lumbar degenerative disease from July 2009 to June 2012. There were 38 cases with whole spine plates and 88 cases with lumbar plates. All of them received magnetic resonance imaging (MRI) scans. Through software Image J, the following spino-pelvic sagittal balance parameters were measured: sagittal vertical axis (SVA, distance between C7 plumb line and posterior upper corner of S1 endplate), thoracic kyphosis (TK, T5-T12 Cobb angle), thoracolumbar kyphosis (TLK, T10-L2 Cobb angle), lumbar lordosis (LL, L1-L5 Cobb angle), pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), T1 spino-pelvic inclination (T1-SPI), T9 spino-pelvic inclination (T9-SPI), spino-sacral angle (SSA), spino-pelvic angle (SPA) and C7 tilt (C7T). Based on the grading system of Pfirrmann et al, the intervertebral disc degeneration grades were acquired for each lumbar segment. According to the normal range of previous studies, each spino-pelvic sagittal balance parameter was classified into 3 groups, i.e. Group A (less than normal), Group B (normal) and Group C (more than normal). All statistical analyses were performed to compare the differences of each lumbar segment/intervertebral disc degeneration among groups by one-way ANOVA test via SPSS 18.0. And P-value < 0.05 was deemed significant. RESULTS: PI had a significant impact on the L5/S1 disc degeneration. And L5/S1 disc degeneration had a significant impact on sagittal balance. Among 57 cases of large PI, LL, PT, SS, SVA, C7T, SPA in L5/S1 of mild-to-moderate and severe degeneration groups was 36.91 ± 14.93 and 29.71 ± 11.30 (P = 0.045), 18.82 ± 8.27 and 25.18 ± 10.19 (P = 0.012), 39.45 ± 8.82 and 34.57 ± 8.88 (P = 0.042), 23.04 ± 26.63 and 62.15 ± 33.82 (P = 0.002), 88.85 ± 3.13 and 83.98 ± 4.62 (P = 0.003), 157.88 ± 11.20 and 147.75 ± 13.98 (P = 0.043) respectively. TLK in L1/L2 with mild-to-moderate and severe degeneration groups was 4.59 ± 7.81 and 14.91 ± 14.75 (P = 0.026). All of L1/L2, L2/L3 and L3/L4 disc degeneration had a significant impact on SPA. SPA in L1/L2, L2/L3, and L3/L4 with mild-to-moderate and severe degeneration groups was 158.47 ± 11.97 and 147.84 ± 12.72 (P = 0.031), 159.91 ± 10.78 and 148.75 ± 13.17 (P = 0.024), 158.46 ± 10.86 and 148.61 ± 13.93 (P = 0.047) respectively. CONCLUSION: With an important impact on lumbar disc degeneration, PI, either too big or too small, may predispose to the occurrences of lumbar disc degeneration. L5/S1 disc degeneration has a significant impact on pelvis postural parameters (PT, SS). L5/S1 degeneration is a key causative factor of pelvic posterior rotation and compensatory process. L5/S1 disc degeneration has an important impact on pelvis overall parameters (SVA, SPA, C7T). And the degeneration of L5/S1 is a key cause of trunk imbalance.

ConvWin-UNet: UNet-like hierarchical vision Transformer combined with convolution for medical image segmentation.

Convolutional Neural Network (CNN) plays a vital role in the development of computer vision applications. The depth neural network composed of U-shaped structures and jump connections is widely used in various medical image tasks. Recently, based on the self-attention mechanism, the Transformer structure has made great progress and tends to replace CNN, and it has great advantages in understanding global information. In this paper, the ConvWin Transformer structure is proposed, which refers to the W-MSA structure in Swin and combines with the convolution. It can not only accelerate the convergence speed, but also enrich the information exchange between patches and improve the understanding of local information. Then, it is integrated with UNet, a U-shaped architecture commonly used in medical image segmentation, to form a structure called ConvWin-UNet. Meanwhile, this paper improves the patch expanding layer to perform the upsampling operation. The experimental results on the Hubmap datasets and synapse multi-organ segmentation dataset indicate that the proposed ConvWin-UNet structure achieves excellent results. Partial code and models of this work are available at https://github.com/xmFeng-hdu/ConvWin-UNet.

Correlation between health literacy and life quality in elderly patients with chronic periodontitis.

OBJECTIVES: This study aims to explore the status quo of health literacy and quality of life in elderly patients with chronic periodontitis and determine their correlation to understand the influencing factors of health literacy and quality of life. METHODS: A total of 150 elderly patients with chronic periodontitis were investigated using the general information questionnaire, Oral Health Literacy-Adult Questionnaire, and Oral Health Impact Profile for Chronic Periodontitis. RESULTS: The total scores of health literacy and quality of life of elderly patients with chronic periodontitis were 6.59±4.11 and 29.25±9.42, respectively. The total score of health literacy was negatively correlated with the total score of quality of life (r=-0.234, P<0.01). Regression analysis showed that age, place of residence, education level, and disease understanding were the influencing factors of health literacy. Family history, smoking, and education level were the influencing factors of quality of life. CONCLUSIONS: The health literacy level of elderly patients with chronic periodontitis is insufficient, and their quality of life needs to be improved. Improving the health literacy of patients is an effective way to improve their quality of life. Nursing staff must formulate corresponding health education strategies to improve the health literacy of patients and improve their quality of life.

Maternal effects drive intestinal development beginning in the embryonic period on the basis of maternal immune and microbial transfer in chickens.

BACKGROUND: Nutrition drives immunity and health in animals, and maternal immunity benefits offspring. In our previous study, a nutritional intervention strategy was found to promote the immunity of hens, which subsequently improved immunity and growth in offspring chicks. Maternal effects clearly exist, but how are mothers' immune advantages transferred to their offspring, and how do they benefit them? RESULTS: Here, we traced the beneficial effects back to the process of egg formation in the reproductive system, and we focused on the embryonic intestinal transcriptome and development, as well as on maternal microbial transfer in offspring. We found that maternal nutritional intervention benefits maternal immunity, egg hatching, and offspring growth. The results of protein and gene quantitative assays showed that the transfer of immune factors into egg whites and yolks depends on maternal levels. Histological observations indicated that the promotion of offspring intestinal development begins in the embryonic period. Microbiota analyses suggested that maternal microbes transfer to the embryonic gut from the magnum to the egg white. Transcriptome analyses revealed that offspring embryonic intestinal transcriptome shifts are related to development and immunity. Moreover, correlation analyses showed that the embryonic gut microbiota is correlated with the intestinal transcriptome and development. CONCLUSIONS: This study suggests that maternal immunity positively influences offspring intestinal immunity establishment and intestinal development beginning in the embryonic period. Adaptive maternal effects might be accomplished via the transfer of relatively large amounts of maternal immune factors and by shaping of the reproductive system microbiota by strong maternal immunity. Moreover, reproductive system microbes may be useful resources for the promotion of animal health. Video Abstract.

[Correlation of adult spinal sagittal imbalance and life quality].

OBJECTIVE: To evaluate the correlation of spinal sagittal imbalance and life quality. METHODS: Radiographic analysis for 48 consecutive symptomatic patients with spinal sagittal imbalance was performed with posteroanterior and lateral standing radiographs. There were 12 males and 36 females with an average age of (66.2 ± 8.5) yrs. The measurement parameters included C7PL, thoracic kyphosis (TK), thoracolumbar kyphosis (TLK), lumbar lordosis (LL), pelvic tilt (PT), pelvic incidence (PI) and sacral slope (SS). Life quality was assessed with SF-36 questionnaire. Pearman's method was employed to analyze the correlation. RESULTS: Mean C7PL was (44.7 ± 22.5) mm, TK (26.1 ± 13.1)°, TLK (11.9 ± 10.3)°, LL (23.5 ± 18.2)°, PT (32.1 ± 13.4)°, PI (57.4 ± 10.9)° and SS (22.5 ± 11.5)°. C7PL had a significant correlation with physical functioning (r = -0.428, P < 0.01) and general health (r = -0.428, P < 0.01). PI had a significant correlation with bodily pain (r = -0.374, P < 0.01), vitality (r = -0.303, P < 0.01), social functioning (r = -0.309, P < 0.01) and role emotional (r = -0.429, P < 0.05). TK had a significant correlation with physical functioning (r = -0.292, P < 0.05) and general health (r = -0.389, P < 0.01). LL had a significant correlation with physical functioning (r = 0.428, P < 0.01), general health (r = 0.340, P < 0.05) and vitality (r = 0.373, P < 0.01). PT had significant correlation with vitality (r = -0.385, P < 0.01) and social functioning (r = -0.417, P < 0.05). No significant correlation existed between TLK, SS and SF-36 categories. CONCLUSION: C7PL, TK, LL, PI and PT are significant parameters correlating with quality of life. PI is the most important one affecting bodily pain. TK, LL and C7PL are the main parameters affecting general health. PI, PT and LL affect vitality the most. Correcting these parameters while treating sagittal imbalance is important for a better life quality.

Association between Klebsiella pneumoniae and ankylosing spondylitis: A systematic review and meta-analysis.

AIM: The aim of this study is to evaluate the association between Klebsiella pneumoniae infection and ankylosing spondylitis (AS). METHOD: Five electronic databases, PubMed, Embase, Medline, Web of Science, and Scopes, were searched until September 29, 2021. Cohort and case-control studies that assessed the association between K. pneumoniae infection and AS were included. Pooled odds ratio (OR) was selected to show the effect size. Subgroup analysis (active or inactive AS) and 2 forms of sensitivity analysis were conducted. All statistical analyses were conducted by using STATA 12.0. RESULTS: There were 25 case-control studies finally included, including 8 studies concerning presence of K. pneumoniae in feces, and 17 studies concerning serum antibody (immunoglobulin [Ig]G, IgM, IgA) against K. pneumoniae. The results suggested that when compared with healthy people, presence of K. pneumoniae in feces was associated with AS (OR: 5.65; 95% CI: 1.68-19.00). Similarly, when compared with healthy people, higher positive rates of IgA (OR: 6.28; 95% CI: 3.32-11.91) and IgG (OR: 5.22; 95% CI: 1.36-19.99) were observed. Subgroup analyses suggested that association between K. pneumoniae and AS appears stronger in active AS. CONCLUSION: When compared with healthy people, a significantly higher positive rate of K. pneumoniae in feces, serum IgA and IgG were observed in patients with AS, suggesting that K. pneumoniae probably plays a crucial role in the occurrence of AS. The findings in this study need further prospective investigations for confirmation.

Wnt and Smad signaling pathways synergistically regulated the osteogenic differentiation of fibroblasts in ankylosing spondylitis.

This study aimed to reveal the cross association of the Wnt-Smad pathway in AS fibroblasts. Fibroblasts from hip synovial tissue were isolated, and the expressions of Wnt and Smad pathway proteins were detected by western blot analysis. DKK-1 and SIS3 were used to inhibit Wnt and Smad, respectively. The proliferation and apoptosis of AS fibroblasts were detected by CCK-8 and flow cytometry, respectively. And the effects of DKK-1 and/or SIS3 on osteogenic differentiation of AS fibroblasts were investigated by alkaline phosphatase (ALP) activity, intracellular calcium concentration, mineralization and osteogenic proteins expressions. Results showed that AS fibroblasts had stronger osteogenic differentiation ability, accompanied by activation of Wnt and Smad pathways. In AS fibroblasts, inhibition of Wnt by DKK-1 could affect the Smad pathway, while inhibition of Smad by SIS3 affect the Wnt pathway, DKK-1 and SIS3 showed a synergistic effect on the regulation of Wnt and Smad. In addition, DKK-1 and SIS3 also showed synergistic positive effects in inhibiting the proliferation and osteogenic differentiation of AS fibroblasts. In conclusion, there was an interaction between Wnt and Smad pathways in AS fibroblasts. DKK-1 and SIS3 can synergistically and negatively regulate the osteogenic differentiation of AS fibroblasts, showing positive effects on AS.

HDAC1 regulates inflammation and osteogenic differentiation of ankylosing spondylitis fibroblasts through the Wnt-Smad signaling pathway.

Ankylosing spondylitis (AS) is a refractory autoimmune disease, whose typical pathology is the development of inflammation to ossification and ankylosis. Histone deacetylase 1 (HDAC1) is considered to be a key factor involved in inflammatory gene transduction, but its role in AS remains unclear. The purpose of this study was to explore the role and possible mechanism of HDAC1 in AS based on the Wnt-Smad pathway. Fibroblasts were isolated from hip synovial tissues of AS patients, adeno-associated virus (AAV) was used to regulate the expression of HDAC1, DKK-1 and SIS3 was used to inhibit Wnt and Smad, respectively. The expressions of Wnt-Smad pathway-related proteins were analyzed by WB, and the TRP ion channel proteins were analyzed by immunofluorescence and WB. The proliferation of AS fibroblasts was detected by CCK-8, the expression of inflammatory cytokines was detected by ELISA, and the effects of HDAC1 on osteogenic differentiation of AS fibroblasts were investigated by alkaline phosphatase (ALP) activity, intracellular calcium concentration, mineralization and osteogenic proteins expressions. Results showed that HDAC1 significantly affected the protein expressions of the Wnt-Smad pathway in AS fibroblasts, and Wnt inhibitor DKK-1 and Smad3 inhibitor SIS3 could significantly reverse the effect of HDAC1 on the Wnt-Smad pathway. In addition, HDAC1 significantly activated the TRP ion channel and promoted the proliferation, inflammatory response and osteogenic differentiation of AS fibroblasts. DKK-1 or SIS3 treatment significantly inhibit the effect of HDAC-1 on AS fibroblasts, suggesting that the Wnt-Smad pathway is involved in the regulation of AS by HDAC1. In conclusion, HDAC1 promotes the proliferation, inflammatory response and osteogenic differentiation of AS fibroblasts through the Wnt-Smad pathway.

Role of death receptor, mitochondrial and endoplasmic reticulum pathways in different stages of degenerative human lumbar disc.

Intervertebral disc (IVD) cell apoptosis has been suggested to play an important role in promoting the degeneration process. It has been demonstrated that IVD cell apoptosis occurs through either death receptor, mitochondrial or endoplasmic reticulum (ER) pathway. Our study aimed to explore the relationship among these three pathways and grade of IVD degeneration (IVDD). IVDs were collected from patients with lumbar fracture, vertebral tumor, disc herniation or spondylolisthesis. IVDs were distinguished by MRI and histomorphological examination, cell apoptosis was detected by TUNEL staining. Biomarkers of these three apoptosis pathways were detected by RT-PCR and Western blot. Furthermore, the correlation between apoptosis pathways biomarkers and disc pathology were analyzed. Nucleus pulposus cell density decreased with degeneration process, and increased apoptotic ratio. ER pathway was predominant in mild stage of IVDD (GRP78, GADD153 upregulation and caspase-4 activation), death receptor pathway was predominant in mild and moderate stages (Fas, FasL up-regulation and caspase-8 activation) and mitochondrial pathway was predominant in moderate and severe stages (Bcl-2 down-regulation, Bax up-regulation, cytochrome-c accumulation in cytoplasm and caspase-9 activation). There were significant differences in the expressions of Fas, FasL, Bax, GADD153, cytochrome-c and cleaved caspase-8/9/3 between contained and non-contained discs. In conclusion, apoptosis occurs via these three apoptosis pathways together in IVDD. ER pathway plays a more critical role in the mild compared to moderate and severe stages, death receptor pathway in mild and moderate, and mitochondrial pathway in moderate and severe stages of IVDD. Disc cells apoptosis may progress rapidly after herniation, and may depend on the type of herniation.

[Experimental study of percutaneous vertebroplasty with a novel bone void filling container system].

OBJECTIVE: To investigate vertebral augmentation with a novel reticulate bone filling container system by polymethyl methacrylate (PMMA) injection in cadaveric simulated vertebral compressive fracture and explore the effect of reticulate bone filling container on cement distribution controlling within vertebral body and the restoration of biomechanical properties after augmentation. METHODS: A total of 28 freshly frozen human vertebrae specimens were randomly divided into 4 groups. After the measurements of bone mineral density (BMD) and vertebral height, each vertebra received an axle load by a MTS (material testing system) machine to test the initial strength and stiffness. Subsequently a simultaneous compressive fracture model was created to measure the stiffness and height of fractured vertebrae. Then the augmentation procedure was performed. Afterward the biomechanical properties and the vertebral height were similarly measured as pre-operatively. The expansion of bone filling container and the distribution of cement within vertebral body were morphologically observed by crossing the specimens in sagittal midline and also integrated with the radiographic results. RESULTS: Stiffness was significantly restored comparing with that of fractured level (P < 0.05). And the bipedicular groups had better restoration results than the unipedicular groups. The strength and height of specimens significantly increased after the augmentation procedure but without difference among groups. In axial radiographic view, the distribution of cement in vertebral body was oval or long oval-shaped in double-layer bone filling container groups while it was irregular in single-layer groups. After crossing, the double-layer version expanded well in vertebral body and could enwrap most of injected cement. There was only a little leakage near the vessel layer. But the single-layer version had a poor expansion and a large amount of cement leakage. CONCLUSION: This novel reticulate bone void filling container system with different layers may restore both the biomechanical properties and the height of fractured vertebrae. But, with the benefit of reducing cement leakage, a double-layer design can enwrap most of injected PMMA and has a brighter prospect of clinical application.