PSD95 as a New Potential Therapeutic Target of Osteoarthritis: A Study of the Identification of Hub Genes through Self-Contrast Model.

Osteoarthritis (OA) is a worldwide joint disease. However, the precise mechanism causing OA remains unclear. Our primary aim was to identify vital biomarkers associated with the mechano-inflammatory aspect of OA, providing potential diagnostic and therapeutic targets for OA. Thirty OA patients who underwent total knee arthroplasty were recruited, and cartilage samples were obtained from both the lateral tibial plateau (LTP) and medial tibial plateau (MTP). GO and KEGG enrichment analyses were performed, and the protein-protein interaction (PPI) assessment was conducted for hub genes. The effect of PSD95 inhibition on cartilage degeneration was also conducted and analyzed. A total of 1247 upregulated and 244 downregulated DEGs were identified. Significant differences were observed between MTP and LTP in mechanical stress-related genes and activated sensory neurons based on a self-contrast model of human knee OA. Cluster analysis identified DLG4 as the hub gene. Cyclic loading stress increased PSD95 (encoded by DLG4) expression in LTP cartilage, and PSD95 inhibitors could alleviate OA progression. This study suggests that inhibiting PSD95 could be a potential therapeutic strategy for preventing articular cartilage degradation.

Nerve Growth Factor-Preconditioned Mesenchymal Stem Cell-Derived Exosome-Functionalized 3D-Printed Hierarchical Porous Scaffolds with Neuro-Promotive Properties for Enhancing Innervated Bone Regeneration.

The essential role of the neural network in enhancing bone regeneration has often been overlooked in biomaterial design, leading to delayed or compromised bone healing. Engineered mesenchymal stem cells (MSCs)-derived exosomes are becoming increasingly recognized as potent cell-free agents for manipulating cellular behavior and improving therapeutic effectiveness. Herein, MSCs are stimulated with nerve growth factor (NGF) to regulate exosomal cargoes to improve neuro-promotive potential and facilitate innervated bone regeneration. In vitro cell experiments showed that the NGF-stimulated MSCs-derived exosomes (N-Exos) obviously improved the cellular function and neurotrophic effects of the neural cells, and consequently, the osteogenic potential of the osteo-reparative cells. Bioinformatic analysis by miRNA sequencing and pathway enrichment revealed that the beneficial effects of N-Exos may partly be ascribed to the NGF-elicited multicomponent exosomal miRNAs and the subsequent regulation and activation of the MAPK and PI3K-Akt signaling pathways. On this basis, N-Exos were delivered on the micropores of the 3D-printed hierarchical porous scaffold to accomplish the sustained release profile and extended bioavailability. In a rat model with a distal femoral defect, the N-Exos-functionalized hierarchical porous scaffold significantly induced neurovascular structure formation and innervated bone regeneration. This study provided a feasible strategy to modulate the functional cargoes of MSCs-derived exosomes to acquire desirable neuro-promotive and osteogenic potential. Furthermore, the developed N-Exos-functionalized hierarchical porous scaffold may represent a promising neurovascular-promotive bone reparative scaffold for clinical translation.

3D-printed vascularized biofunctional scaffold for bone regeneration.

3D-printed biofunctional scaffolds have promising applications in bone tissue regeneration. However, the development of bioinks with rapid internal vascularization capabilities and relatively sustained osteoinductive bioactivity is the primary technical challenge. In this work, we added rat platelet-rich plasma (PRP) to a methacrylated gelatin (GelMA)/methacrylated alginate (AlgMA) system, which was further modified by a nanoclay, laponite (Lap). We found that Lap was effective in retarding the release of multiple growth factors from the PRP-GelMA/AlgMA (PRP-GA) hydrogel and sustained the release for up to 2 weeks. Our in vitro studies showed that the PRP-GA@Lap hydrogel significantly promoted the proliferation, migration, and osteogenic differentiation of rat bone marrow mesenchymal stem cells, accelerated the formation of endothelial cell vascular patterns, and promoted macrophage M2 polarization. Furthermore, we printed hydrogel bioink with polycaprolactone (PCL) layer-by-layer to form active bone repair scaffolds and implanted them in subcutaneous and femoral condyle defects in rats. In vivo experiments showed that the PRP-GA@Lap/PCL scaffolds significantly promoted vascular inward growth and enhanced bone regeneration at the defect site. This work suggests that PRP-based 3D-bioprinted vascularized scaffolds will have great potential for clinical translation in the treatment of bone defects.

[Polymorphisms of vascular diseases-related genes in Guangzhou Biobank Cohort Study].

OBJECTIVE: To study the polymorphisms of cerebrovascular and cardiovascular disease genes using Taqman single nucleotide polymorphism (SNP) genotyping kits. METHODS: A total of 2000 subjects were recruited from the Guangzhou Biobank Cohort Study (GBCS), and 15 SNPs were detected using Taqman SNP genotyping kits and an ABI 7900HT real time PCR system. The data were tested for the Hardy-Weinberg equilibrium, and then compared with the data of the Chinese population from the International HapMap Project (HapMap_HCN). RESULTS: (1) All genotype data of the 15 SNPs were consistent with the Hardy-Weinberg rules. (2) The significant differences were observed among two SNPs, rs4220 and rs5368 and the HapMap_HCN (rs4220 28.2% vs 17.8%; chi(2) = 4.891, P = 0.028; rs5368 22.1% vs 32.2%, chi(2) = 5.137, P = 0.024). Comparing other gene bank data, such as AFD-CHN-PANEL, the Allele Frequency Database (ALFRED) and JBIC-allele, it would be most likely that our observations represent differences between the Northern and Southern populations in China. CONCLUSION: Such Biobank study provided a useful platform for the study of the role of genetic and environmental determinants on cerebrovascular and cardiovascular disease.

[Electrocardiogram change of workers after disengaging occupational noise exposure].

OBJECTIVE: To examine electrocardiogram (ECG) change of workers after leaving occupational noise exposure. METHODS: In the first phase of the Guangzhou Biobank Cohort Study, 10413 Guangzhou residents aged 50 years or more received a face-to face interview including noise exposure history, a full medical check-up and laboratory tests in 2003 - 4. ECG examination was carried out using 12-lead MAC-CS ECG machine made in HP Ltd, Shanghai, China. ECG of every subject was independently diagnosed by two doctors who had obtained ECG diagnosis qualification. RESULTS: (1) Among 10413 subjects, 70% were female, and more than half were aged 60-69 years. The mean age and education level in males were higher than those in females. (2) 2119 subjects (21.0%) were previously exposed to noise and were retired now. (3) The prevalence of ECG abnormality was respectively 82.4% and 79.1% in females and males. Rate of ECG abnormality increased with age in both females and males (P = 0.000), and was lower in females with higher education level (P = 0.000). There was not significant difference in ECG abnormality between workers previously exposed or unexposed to noise in both females and males (P > 0.05). 4 Comparing workers previously exposed to those unexposed to noise in both females and males, after adjusting for age, educational level and smoking status, the rates of arrhythmia, axes deviation, atria and ventricle hypertrophy, ST-T change and coronary artery syndrome were not significantly different (P > 0.05). CONCLUSION: Our study showed that the prevalence of ECG abnormality was not significantly increased in workers previously exposed to noise and had left exposure now compared to those never exposed to noise. Because our study was cross section and occupational history was obtained from a questionnaire but was not confirmed by the companies, further study is needed.

[Clinical manifestation and ultrasonic characteristics of five patients with acute arsenic poisoning].

OBJECTIVE: To study the clinical manifestation and ultrasonic characteristics of liver, kidney and heart of five patients with acute arsenic poisoning. METHODS: The activity of serum myocardial enzymes, function of liver and kidney, and urinary As concentrations were measured. HDI 3000 Enhanced, and Toshiba 38A two dimensional ultrasound was used to examine the ultrasonic echogram of heart, liver, kidney of the patients. RESULTS: (1) The arsenic concentrations in the urine (1.9 approximately 15.6 micromol/L) were higher than the normal value (1.17 micromol/L) in these patients (blood dialytic fluid of one patient with anuria was measured); (2) Four of them had increased WBC, or anemia, and abnormal urine routine to various degree; (3) The activities of serum myocardial enzymes (CK, AST, LDH and HBDH) in 4 patients were at least 2 items increased; (4) Serum bilirubin and urea nitrogen in all patients were increased; (5) The ultrasonic echogram of liver and kidney in these 5 patients showed abnormality to various degree, one of them had slight enlargements in left atrium and ventricle as well as a little pericardial fluid. CONCLUSION: The clinical manifestation and ultrasonic characteristics of liver, kidney, and heart were consistent with the pathologic changes in acute arsenic poisoning. Early blood dialysis may reduce visceral damage.

[Acute fluoroacetamide poisoning with main damage to the heart].

OBJECTIVE: To observe the heart damage in 10 patients with acute fluoroacetamide poisoning. METHODS: Monitoring serum activities of myocardial enzymes [creatine kinase (CK), asparate aminotransferase (AST), lactate dehydrogenase(LDH) and hydroxybutyrate dehydrogenase(HBDH)] and recording ECG on these 10 patients were performed during the period of their hospitalization. In the mean while, 24 hour dynamic ECG were also recorded and analysed using GP7000L Holter. RESULTS: (1) Urinary fluorine ion concentrations were increased in 9 patients before therapy and in all these 10 patients during therapeutic period. (2) The activities of serum CK in 2 patients and that of serum HBDH in one patient were increased before therapy. However, the serum activities of one or more than one myocardial enzymes were increased in all these 10 patients during therapeutic period. (3) Four patients had abnormal change of ECG before therapy and 5 patients during therapeutic period. (4) 24 hour dynamic ECG records showed that there were heart electrical alternans in 9 patients. One patient had wandering pacemaker and 6 patients had arythmia. CONCLUSION: Fluoroacetamide may cause obvious heart damage, and also heart electrical alternation.

[Dose-response relationship between smoking status and carotid atherosclerosis].

OBJECTIVE: To examine the dose-response relationship of smoking status with carotid atherosclerosis in 959 relatively healthy Chinese men. METHODS: 959 older Chinese men were selected from Guangzhou Biobank Cohort Study (GBCS) on cardiovascular disease. Personal histories were collected and fasting plasma glucose and lipids, blood pressure, and common carotid artery intima-median thickness (CCA-IMT) were measured. RESULTS: (1) Composition of the cases: 39.1% were non-smokers, 25.7% were former smokers and 35.2% were current smokers. The mean (95% confidence interval) carotid IMT was 0.78 (0.77 - 0.79) mm. 18.4% of the subjects had carotid IMT equal to or thicker than 1.0 mm while 34.1% had carotid plaque. (2) After adjusting for age, sex, physical activity, body mass index, fasting glucose, triglyceride, high-density lipoprotein cholesterol, systolic and diastolic blood pressure, compared to never smokers, current smokers had significantly increased risk for thicker IMT and carotid plaque [odds ratio (OR) = 1.82, 95%CI: 1.30 - 2.55 and OR = 1.95, 95%CI: 1.38 - 2.75, respectively, all P < 0.001]. The risk for thicker IMT and carotid plaque increased with the increasing amount (cigarettes/day) and duration of smoking (years) as well with cigarette pack-years (P for trend all ≤ 0.01). CONCLUSION: An elevated risk with a clear dose-response relationship was found between cigarette smoking and carotid atherosclerosis. Quitting smoking or reducing the amount of smoking may lower the risk of atherosclerosis, preventing and controlling the occurrence of cardiovascular diseases, and reducing the related cardiovascular mortalities.

[Study on the association between gene polymorphism of ENPP1 and left ventricular hypertrophy: a Guangzhou Biobank Cohort Study-CVD].

OBJECTIVE: To examine the association between genetic polymorphism of rs1409181 in ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) and left ventricular hypertrophy (LVH) among older Chinese in Guangzhou. METHODS: 390 subjects aged ≥ 50 years were randomly selected from the Guangzhou Biobank Cohort Study-CVD. Information on personal history, blood pressure, fasting plasma glucose and lipids were collected. Color Doppler ultrasound was used to measure the indicators of LVH, including left ventricular internal diastolic diameter (LVIDD), thickness of the interventricular septum diastolic wall (IVSD) and the posterior wall diastolic diameter (LVPWD). LVIDD was calculated using Devereux ventricular mass (LVM) equation while the Left ventricular mass index (LVMI) equation was used to estimate LVH. The genotype of rs1409181 was determined by Taqman SNP genotyping kits using the ABI 7900HT real time PCR system. RESULTS: In the GG, CG and CC genotype groups, the proportions of LVH were 21.5%, 28.2% and 37.5% respectively. Compared with GG, the adjusted odds ratios (95% confidence interval) for the LVH were 1.39 (0.78 - 2.50) and 2.36 (1.21 - 4.60) for CG genotype and CC genotype of ENPP1 respectively (P for trend = 0.01). CONCLUSION: Polymorphism of ENPP1 gene rs1409181 was associated with LVH in the older Chinese people in Guangzhou.

[Effect of physical activity strength on the diabetes mellitus prevalence in the elderly under the influence of International Physical Activity Questionnaire].

OBJECTIVE: To examine the impact of International Physical Activity Questionnaire (IPAQ) physical activity intensity on to the prevalence of diabetes mellitus among Chinese elderly. METHODS: A total number of 1996 residents aged 50 or above living in Guangzhou city were recruited from the phase 3 of the Guangzhou Biobank Cohort Study. Information on physical activity and fasting plasma glucose status was derived from standardized interviews and laboratory assays. RESULTS: Among the participants who were classified as physically active (60.0%), moderate active (29.8%) and inactive (10.2%), the prevalence rates of type 2 diabetes mellitus were 9.1%, 12.0% and 14.2%, respectively. After adjustment on age, sex, obesity and other potential confounding factors, data from logistic regression model showed that the odds ratios (95% confidence interval) for diabetes on subjects in physically moderate active and active group were 0.75(0.46-1.26)and 0.60(0.38-0.97) respectively with P for trend as 0.03, when comparing to those physically inactive ones. CONCLUSION: Promotion of physical activity might have had some effects in reducing the risk of diabetes mellitus among the older adults.