Naringenin induces neuroprotection against homocysteine-induced PC12 cells via the upregulation of superoxide dismutase 1 expression by decreasing miR-224-3p expression.

Naringenin is a flavonoid compound with antioxidant effects. It is used to treat oxidative stress-related diseases, but its mechanism is unclear. In this experiment, we explored whether naringenin can increase the expression of superoxide dismutase 1(SOD1), reduce the oxidative stress of PC12 cells induced by homocysteine (Hcy), and decrease the apoptosis of PC12 cells induced by Hcy by inhibiting the expression of mir-224-3p. Different concentrations of Hcy (1, 3, 5, 8, and 10 mmol/L) was used to analyze effect of homocysteine on PC12 cells. A total of 5 mmol/L Hcy was used to induce the excitatory and neurotoxicity model of PC12 cells in vitro. The cells were divided into normal control, Hcy induction, Hcy + Naringenin (25 µM), Hcy + Naringenin (50 µM), Hcy + Naringenin (75 µM), Hcy + Naringenin (100 µM), and Hcy + Naringenin (150 µM) groups. The relative survival rate and activities of the PC12 cells were determined by the MTT method, and the apoptosis rate of the PC12 cells was determined by using flow cytometry. The Western blot method was used to determine the expressions of SOD1, Bax, Caspase-3, Caspase-8, and Bcl-2 in the PC12 cells induced by Hcy. The expressions of SOD1 mRNA and miR-224-3p in the Hcy-induced PC12 cells were determined by RT-PCR. Results found that Hcy increased the expression of miR-224-3p in a dose-dependent manner but decreased that of SOD1 mRNA and protein. Hcy also increased oxidative stress in the PC12 cells and the proapoptotic proteins Bax, Caspase-3, and Caspase-9. Furthermore, it decreased the expression of anti-apoptotic protein Bcl-2 and the activity and survival rate of the HT22 cells, but it increased the apoptosis of the PC12 cells. The treatment of Hcy-induced PC12 cells with different concentrations of naringenin for 24 h decreased the expression of miR-224-3p in a dose-dependent manner and increased the expressions of SOD1 mRNA and protein. The treatment also decreased the oxidative stress in the PC12 cells and the expressions of pro-apoptotic proteins Bax, Caspase-3, and Caspase-9; increased the expression of anti-apoptotic protein Bcl- 2; decreased the apoptosis of the PC12 cells; and increased the PC12 cells.The results suggest that Naringenin can decrease the apoptosis and oxidative stress of PC12 cells induced by Hcy and increase the activities and survival rates of PC12 cells. The mechanism may be related to naringenin decreasing the expression of miR-224-3p in PC12 cells induced by Hcy and increasing the expressions of SOD1 mRNA and protein.

Impact of individual behaviour change on the spread of emerging infectious diseases.

Human behaviour plays an important role in the spread of emerging infectious diseases, and understanding the influence of behaviour changes on epidemics can be key to improving control efforts. However, how the dynamics of individual behaviour changes affects the development of emerging infectious disease is a key public health issue. To develop different formula for individual behaviour change and introduce how to embed it into a dynamic model of infectious diseases, we choose A/H1N1 and Ebola as typical examples, combined with the epidemic reported cases and media related news reports. Thus, the logistic model with the health belief model is used to determine behaviour decisions through the health belief model constructs. Furthermore, we propose 4 candidate infectious disease models without and with individual behaviour change and use approximate Bayesian computation based on sequential Monte Carlo method for model selection. The main results indicate that the classical compartment model without behaviour change and the model with average rate of behaviour change depicted by an exponential function could fit the observed data best. The results provide a new way on how to choose an infectious disease model to predict the disease prevalence trend or to evaluate the influence of intervention measures on disease control. However, sensitivity analyses indicate that the accumulated number of hospital notifications and deaths could be largely reduced as the rate of behaviour change increases. Therefore, in terms of mitigating emerging infectious diseases, both media publicity focused on how to guide people's behaviour change and positive responses of individuals are critical.

Diabetes incidence and prevalence in Hong Kong, China during 2006-2014.

AIMS: To estimate recent secular changes in the incidence and prevalence of diabetes and pre-diabetes among Hong Kong Chinese adults, and thus show possible future trends for developing mainland China. METHODS: Based on a complete census of the public sector health records of 6.4 million people from 2006 to 2014, diabetes cases were ascertained using different methods including the World Health Organization (WHO) 2011 guidelines (HbA1c , fasting plasma glucose and glucose tolerance test), American Diabetes Association (ADA) 2015 guidelines (plus random plasma glucose), and additionally recorded diagnosis codes and medication dispensation. Pre-diabetes was defined using ADA 2015 guidelines. RESULTS: We identified 697 201 people with diabetes (54.2% were incident cases); and 1 229 731 people with diabetes or pre-diabetes. In 2014, the overall incidence of diabetes was 9.46 per 1000 person-years [95% confidence interval (CI): 9.38 to 9.54], and overall prevalence was 10.29% (95% CI: 10.27% to 10.32%). Incidence of diabetes decreased significantly from 2007 to 2014 (quadratic trend, P < 0.001). From 2006 to 2014, the prevalence of diabetes increased significantly in both sexes and across all age groups (quadratic trend, P < 0.001). The overall incidence of pre-diabetes in 2014 was 18.88 per 1000 person-years (95% CI: 18.76 to 18.99), and the overall prevalence of pre-diabetes was 8.90% (95% CI: 8.87% to 8.92%). CONCLUSIONS: Similar to other developed western and Asian populations, diabetes (and pre-diabetes) incidence in Hong Kong Chinese appeared to have stabilized and there have been small declines during the period of observation. Ageing and survivorship will likely drive a continued increase in the prevalence of diabetes and pre-diabetes, albeit with a decelerating growth rate if past trends persist.

[Effect of simulating leg length inequality on spinal and pelvic posture in the elderly].

Objective: To study the effects of simulating leg length inequality on the spine and pelvic posture in standing and walking states and to explore their compensatory laws. Methods: From January to April, a total of 44 healthy volunteers were rasterstereographically examined for spine and pelvis in Institute of Digitized Medicine, Wenzhou Medical University and Department of Orthopaedics, First Affiliated Hospital of Wenzhou Medical University.Volunteers wore uniform shoes, and single 5 mm thick insoles were customized.The simulating leg length inequalities (5-30 mm) were artificially created by increasing insole height.The parameters of 3D body surface parameters and 4D dynamic parameters of the pelvic and spine were measured and statistically analyzed in standing and walking states. Results: In the static standing state, with the increase of the difference of both lower extremities, coronal plane pelvic tilt and sagittal plane pelvic torsion also increased[the maximum value about (10.6±4.3) mm and (3.3±3.5)°], as well as the frontal deviation of the spine [the maximum value about (11.1±17.9) mm]. But the pelvic rotation, vertebral surface rotation angle (rms) and spine sagittal plane deviation were no obvious changes.In the walking state, with the difference between lower extremities increased, the maximum angles of vertebral surface rotation to the left and right and pelvic rotation to the left and right were no obvious changes, but (coronal) spinal maximum offset distance to left and right increased [the maximum value about (9.8±5.1), (10.4±6.9) mm]. Conclusion: The effect of the leg length discrepancy on the pelvic coronal plane and the sagittal plane changes are obvious, but little effect has on the pelvic cross section.The pelvis is compensated by the increase of the inclination of the coronal plane and the sagittal angle at first order.Similarly, the effect on the coronal plane of the spine is more markedly, but the changes of sagittal and cross-section of the spine is less affected, the spine is mainly compensated by the coronal plane bending at second order.

Prediction of en-route complications during interfacility transport by outcome predictive scores in ED.

OBJECTIVE: The objective was to determine the accuracy of the outcome predictive scores (Modified Early Warning Score [MEWS]; Hypotension, Low Oxygen Saturation, Low Temperature, Abnormal ECG, Loss of Independence [HOTEL] score; and Simple Clinical Score [SCS]) in predicting en-route complications during interfacility transport (IFT) in emergency department. DESIGN: This was a retrospective cohort study. METHODS: All IFT cases by ambulances with either nurse-led or physician-led escort, occurring between 1 January 2011 and 31 December 2012, were included. Obstetric and pediatric cases (age < 18 years) were excluded. The condition of patients was quantified by using the predictive scores (MEWS, HOTEL, and SCS) at triage station and on ambulance departure. The accuracy of predictive scores was compared by the receiver operating characteristic (ROC) curves. RESULTS: A total of 659 cases were included. Seventeen cases had en-route complications (2.6%). The complication rate in physician-escorted transport (2.2%) was similar to that in nurse-escorted transport (2.6%). None of the 57 intubated cases had en-route complications. The area under the ROC curve for MEWS was 0.662 (triage) and 0.479 (departure). The accuracy of MEWS at triage was better than that at departure (P = .049). The area under the ROC curve for HOTEL was 0.613 (triage) and 0.597 (departure), and that for SCS was 0.6 (triage) and 0.568 (departure). In general, the predictive scores at triage were better than those on departure. CONCLUSION: None of the scores had good accuracy in prediction of en-route complications during IFT. MEWS at triage was among the best one already but was not ideal.

Hypoxia facilitates epithelial-mesenchymal transition-mediated rectal cancer progress.

Rectal cancer is a commonly observed tumor in clinics, and epithelial-mesenchymal transition (EMT) is very important for tumor invasion and metastasis. We established a rectal cancer HCT-116 cell hypoxia model and detected cell proliferation, invasion, and EMT-related protein expression in this model, aiming to analyze the effect of hypoxia on rectal cancer cell EMT. Rectal cancer cell line HCT-116 was cultured in normoxic, hypoxic, or anaerobic environment, and hypoxia-inducible factor-1α (HIF-1α) mRNA expression was detected in the cells by real-time PCR. Cell proliferation was tested by MTT assay; cell invasion was determined by transwell assay, and HIF-1α, epithelial-cadherin, and Snail protein levels were evaluated by western blot analysis. HIF-1α mRNA level significantly increased in the anaerobic group compared to that in the normoxic and hypoxic groups (P < 0.05). HCT-116 cell proliferation in the anaerobic group was obviously higher than that in the other two groups, with the hypoxic group showing stronger proliferative ability than the normoxic group (P < 0.05). Compared to the normoxic group, the HCT-116 cells demonstrated enhanced cell invasion and migration in hypoxic and anaerobic groups. HIF-1α and Snail expressions were upregulated, whereas epithelial-cadherin expression had declined in the hypoxic and anaerobic groups, compared to those in the normal control (P < 0.05). Therefore, hypoxia promoted rectal cancer cell progress by increasing HIF-1α to induce EMT.

[Interaction of DAXX and human papillomavirus type 16 E2 protein].

The aim of the study was to explore the interactions of human papilloma virus 16 (HPV16) E2 protein and Daxx. The location or co-localization of PML and E2 with Daxx in Caski cells was observed by indirect immunofluorescence test. The interaction of E2 and Daxx was analyzed by co-immunoprecipitation, Western-blot and yeast-two hybrid assay. In Caski cells the fluorescence of Daxx or PML was mainly distributed in the cytoplasm or nucleus, respectively, and in the align image their signals did not overlapped. However, when the red signal of HPV16 E2 and the green signal of Daxx in cyto- plasm of Caski cells were merged, the yellow signals appeared. The yeast co-transformed with pGBKT7/Daxx and pGADT7/E2 or pGADT7/E2 TAD can grow onto SD/-Trp-Leu-His and SD/-Trp-Leu-His-Ade plates. So Daxx wasn't co-located with PML but with HPV16 E2 mainly in the cytoplasm of Caski cells. On the base of the results one can propose that HPV16 E2, in particularly its transcription-activity domain (TAD), interacts with Daxx.

Differences in non-enzymatic glycation and collagen cross-links between human cortical and cancellous bone.

UNLABELLED: It is important to establish the relationship between pentosidine and advanced glycation endproducts (AGEs) in bone. We found the relationship between pentosidine and AGEs and their magnitude of accumulation were dependent on bone's surface-to-volume ratio. Results illustrate the importance of measuring pentosidine and AGEs separately in cancellous and cortical bone. INTRODUCTION: Accumulation of collagen cross-links (AGEs) produced by non-enzymatic glycation deteriorates bone's mechanical properties and fracture resistance. Although a single AGE, pentosidine, is commonly used as a representative marker, it is unclear whether it quantitatively reflects total fluorescent AGEs in bone. The goal of this study was to establish the relationship between pentosidine and total AGEs in cancellous and cortical bone. METHODS: Pentosidine and total AGEs were quantified in 170 human bone samples. Total fluorescent AGEs were measured in 28 additional cancellous and cortical bone specimens of the same apparent volume that were incubated in control or in vitro glycation solutions. Correlations between pentosidine and total AGEs and differences between cortical and cancellous groups were determined. RESULTS: Pentosidine was correlated with total AGEs in cancellous bone (r = 0.53, p < 0.0001) and weakly correlated in cortical bone (r = 0.23, p < 0.05). There was more pentosidine (p < 0.01) and total AGEs (p < 0.001) in cancellous than in cortical bone. The in vitro glycation substudy showed that cancellous bone accumulated more AGEs than cortical bone (p < 0.05). CONCLUSION: The relationship between pentosidine and total AGEs and their magnitude of accumulation differed in cancellous and cortical bone of the same apparent volume, and were dependent on the surface-to-volume ratios of each sample. It is important to consider the bone types as two separate entities, and it is crucial to quantify total AGEs in addition to pentosidine to allow for more comprehensive analysis of the effects of non-enzymatic glycation in bone.

A critical review on the current state of antimicrobial glove technologies: advances, challenges and future prospects.

Following recent viral outbreaks, there has been a significant increase in global demand for gloves. Biomedical research focuses increasingly on antimicrobial gloves to combat microbial transmission and hospital-acquired infections. Most antimicrobial gloves are manufactured using antimicrobial chemicals such as disinfectants, biocides and sanitizers. The design of antimicrobial gloves incorporates advanced technologies, including colloidal particles and nanomaterials, to enhance antimicrobial effectiveness. A category of antimicrobial gloves also explores and integrates natural antimicrobial benefits from animals, plants and micro-organisms. Many types of antimicrobial agents are available; however, it is crucial that the selected agent exhibits a broad spectrum of activity and is not susceptible to promoting resistance. Additionally, future research should focus on the potential effect of antimicrobial gloves on the skin microbiota and irritation during extended wear. Careful integration of the antimicrobial agent is essential to ensure optimal effectiveness without compromising the mechanical properties of the gloves.

Omentin-1 exerts bone-sparing effect in ovariectomized mice.

UNLABELLED: Omentin-1 inhibited osteoblast differentiation in vitro. In co-culture systems of osteoblasts and osteoclast precursors, omentin-1 reduced osteoclast formation by stimulating osteoprotegerin (OPG) and inhibiting receptor activator for nuclear factor κB ligand (RANKL) production in osteoblasts. In vivo, adenovirus-mediated overexpression of omentin-1 suppressed bone turnover and restored bone mineral density (BMD) and bone strength in ovariectomized mice. INTRODUCTION: Omentin-1 (also intelectin-1) is a recently identified visceral adipose tissue-derived cytokine that is highly abundant in plasma. This study was undertaken to investigate the effects of omentin-1 on bone metabolism. METHODS: Osteoblast differentiation was assessed by measuring alkaline phosphatase activity, osteocalcin production and matrix mineralization. OPG and RANKL protein expression and secretion in osteoblasts were detected by Western blot and ELISA, respectively. The effect of recombinant omentin-1 on osteoclast formation was examined in co-culture systems of osteoblasts and osteoclast precursors. The effects of intravenous administration of adenoviral-delivered omentin-1 on bone mass, bone strength, and bone turnover were also examined in ovariectomized mice. RESULTS: In vitro, omentin-1 inhibited osteoblast differentiation, while it had no direct effect on osteoclast differentiation; it also reduced osteoclast formation in the co-culture systems through stimulating OPG and inhibiting RANKL production in osteoblasts. In vivo, adenovirus-mediated overexpression of omentin-1 partially restored BMD and bone strength in ovariectomized mice, accompanied by decreased levels of plasma osteocalcin and tartrate-resistant acid phosphatase-5b and lower serum RANKL/OPG ratios. CONCLUSION: The present study suggests that omentin-1 ameliorates bone loss induced by estrogen deficiency via downregulating the RANKL/OPG ratio.

The effect of dichloroacetate on health- and lifespan in C. elegans.

Aging is associated with increased vulnerability to chronic, degenerative diseases and death. Strategies for promoting healthspan without necessarily affecting lifespan or aging rate have gained much interest. The mitochondrial free radical theory of aging suggests that mitochondria and, in particular, age-dependent mitochondrial decline play a central role in aging, making compounds that affect mitochondrial function a possible strategy for the modulation of healthspan and possibly the aging rate. Here we tested such a "metabolic tuning" approach in nematodes using the mitochondrial modulator dichloroacetate (DCA). We explored DCA as a proof-of-principle compound to alter mitochondrial parameters in wild-type animals and tested whether this approach is suitable for reducing reactive oxygen species (ROS) production and for improving organismal health- and lifespan. In parallel, we addressed the potential problem of operator bias by running both unblinded and blinded lifespan studies. We found that DCA treatment (1) increased ATP levels without elevating oxidative protein damage and (2) reduced ROS production in adult C. elegans. DCA treatment also significantly prolonged nematode health- and lifespan, but did not strongly impact mortality doubling time. Operator blinding resulted in considerably smaller lifespan-extending effects of DCA. Our data illustrate the promise of a "metabolic tuning" intervention strategy, emphasize the importance of mitochondria in nematode aging and highlight operator bias as a potential confounder in lifespan studies.

Resource implications of inter-facility transport between emergency departments in Hong Kong.

BACKGROUND: The Inter-facility Transport (IFT) service provided by the Emergency Department (ED) is a vital service in Hong Kong. Patients need to be rapidly transported over distances to access appropriate healthcare facilities. METHODS: This study aims 1. to examine the resource utilisation of IFT accompanied by ED staff and 2. to analyse the crude, fixed and variable costs of IFT. A retrospective review was conducted of all IFT from Alice Ho Miu Ling Nethersole Hospital in the New Territories of Hong Kong where ED staff accompanied patients from 1 January 2006 to 31 December 2008. Descriptive analysis was used to evaluate the crude, fixed and variable costs per year for providing an ED-based IFT service. RESULTS: There were 337 transports accompanied by either medical or nursing staff from the ED that accounted for around 2% of all IFT. The most common indication for mobilising the transport team was an unstable clinical condition that required neurosurgical care. The average transport service time was 57.7 min per transport (SD 11.0). Resource utilisation consisted of fixed and variable costs that summed up to a cost of HKD $87,224.3 (USD $11,182.6) per year and the crude cost of providing IFT service by the ED was HKD$852.2 (USD $109.3) per patient. CONCLUSION: The crude cost of providing IFT service by the ED was reasonable and acceptable.

Effect of diltiazem on exercise tolerance in patients with stable coronary artery disease and hypertension.

The present study aimed to investigate the effect of diltiazem on cardiovascular risk and exercise tolerance in patients with stable coronary artery disease and hypertension. From 2016 to 2018, 80 patients with stable coronary artery disease (5 < Gensini score < 20) and hypertension were enrolled into the present study. These patients were randomly divided into two groups: diltiazem group (Dil, 90 mg, bid), and control group (angiotensin-converting enzyme inhibitors (ACEI)/angiotensin II receptor blockers (ARB) for reducing blood pressure and ß-receptor blockers for controlling heart rate). Liver and kidney function, heart rate variability (HRV), blood pressure variability (BPV) and bicycle exercise were measured at baseline and after six months. The incidence of cardiovascular events (re-hospitalization due to angina pectoris, acute myocardial infarction, and cardiogenic death) was also assessed. The differences in all indexes at baseline between these two groups were not statistically significant (P > 0.05, respectively). After six months of treatment, both groups of patients had significant improvements in HRV, BPV and exercise tolerance compared that before treatment (p < 0.05). The difference in the decrease in systolic blood pressure, improvement of HRV and BPV, and cardiovascular events between these two groups was not statistically significant (P = 0.588, 0.431, 0.152, 1.000, respectively). But the Dil group was significantly better than the control group in degree of heart rate decline, diastolic blood pressure decline, and improvement of ST segment depression (P < 0.001), and the improvement in exercise tolerance was also better than that of the control group. We found that diltiazem compared with ACEI/ARB and ß-receptor blockers can further improve the exercise tolerance of patients with stable coronary artery disease and hypertension.

[Analysis on dynamical mechanism of multi outbreaks of COVID-19].

Objective: In the context of COVID-19 pandemic, the epidemic severities, non-pharmaceutical intervention intensities, individual behavior patterns and vaccination coverage vary with countries in the world. China has experienced a long period without indigenous cases, unfortunately, multi local outbreaks caused by imported cases and other factors have been reported, posing great challenges to COVID-19 prevention and control in China. Thus it is necessary to explore the mechanisms of the re-emerged COVID-19 epidemics and their differences. Methods: Based on susceptible exposed infectious recovered (SEIR) epidemic dynamics model, we developed a set of novel evolution equations which can describe the dynamic processes of integrated influence of interventions, vaccination coverage and individual behavior changes on the re-emergency of COVID-19 epidemic. We developed methods to calculate the optimal intervention intensity and vaccination rate at which the size of susceptible population can be reduced to less than threshold for the re-emergency of COVID-19 epidemic. Results: If strong interventions or super interventions are lifted too early, even a small cause can lead to the re-emergence of COVID-19 epidemic at different degrees. Moreover, the stronger the early control measures lifted are, the more severe the epidemic is. The individual behavior changes for the susceptibility to the epidemic and the enhancement or lifting of prevention and control measures are key factors to influence the incidence the multi outbreaks of COVID-19. The optimist early intervention measures and timely optimization of vaccination can not only prevent the re-emergency of COVID-19 epidemic, but also effectively lower the peak of the first wave of the epidemic and delay its arrival. Conclusion: The study revealed that factors for the re-emergence of COVID-19 epidemics included the intensity and lifting of interventions, the change of individual behavior to the response of the epidemic, external incentives and the transmissibility of COVID-19.

[Prediction modeling with data fusion and prevention strategy analysis for the COVID-19 outbreak].

Since December 2019, the outbreak of COVID-19 in Wuhan has spread rapidly due to population movement during the Spring Festival holidays. Since January 23rd, 2020, the strategies of containment and contact tracing followed by quarantine and isolation has been implemented extensively in mainland China, and the rates of detection and confirmation have been continuously increased, which have effectively suppressed the rapid spread of the epidemic. In the early stage of the outbreak of COVID-19, it is of great practical significance to analyze the transmission risk of the epidemic and evaluate the effectiveness and timeliness of prevention and control strategies by using mathematical models and combining with a small amount of real-time updated multi-source data. On the basis of our previous research, we systematically introduce how to establish the transmission dynamic models in line with current Chinese prevention and control strategies step by step, according to the different epidemic stages and the improvement of the data. By summarized our modelling and assessing ideas, the model formulations vary from autonomous to non-autonomous dynamic systems, the risk assessment index changes from the basic regeneration number to the effective regeneration number, and the epidemic development and assessment evolve from the early SEIHR transmission model-based dynamics to the recent dynamics which are mainly associated with the variation of the isolated and suspected population sizes.

Variability of T2-Relaxation Times of Healthy Lumbar Intervertebral Discs is More Homogeneous within an Individual Than across Healthy Individuals.

BACKGROUND AND PURPOSE: When one uses T2 relaxometry to classify lumbar intervertebral discs as degenerated, it is unclear whether the normative data should be based on other intervertebral discs from the same individual or from a pool of extraneous controls. This study aimed to explore the extent of intra- versus intersubject variation in the T2 times of healthy intervertebral discs. MATERIALS AND METHODS: Using prospectively acquired T2-relaxometry data from 606 intervertebral discs in 101 volunteers without back pain (47 men, 54 women) in a narrow age range (25-35 years), we calculated intra- and intersubject variation in T2 times of intervertebral discs graded by 2 neuroradiologists on the Pfirrmann scale. Intrasubject variation of intervertebral discs was assessed relative to other healthy intervertebral discs (Pfirrmann grade, ≤2) in the same individual. Multiple intersubject variability measures were calculated using healthy extraneous references ranging from a single randomly selected intervertebral disc to all healthy extraneous intervertebral discs, without and with segmental stratification. These variability measures were compared for healthy and degenerated (Pfirrmann grade ≥3) intervertebral discs. RESULTS: The mean T2 values of healthy (493/606, 81.3%) and degenerated intervertebral discs were 121.1 ± 22.5 ms and 91.5 ± 18.6 ms, respectively (P < .001). The mean intrasubject variability for healthy intervertebral discs was 9.8 ± 10.7 ms, lower than all intersubject variability measures (P < .001), and provided the most pronounced separation for healthy and degenerated intervertebral discs. Among intersubject variability measures, using all segment-matched healthy discs as references provided the lowest variability (P < .001). CONCLUSIONS: Normative measures based on the T2 times of healthy intervertebral discs from the same individual are likely to provide the most discriminating means of identifying degenerated intervertebral discs on the basis of T2 relaxometry.

Changes in non-enzymatic glycation and its association with altered mechanical properties following 1-year treatment with risedronate or alendronate.

SUMMARY: One year of high-dose bisphosphonate (BPs) therapy in dogs allowed the increased accumulation of advanced glycation end-products (AGEs) and reduced postyield work-to-fracture of the cortical bone matrix. The increased accumulation of AGEs in these tissues may help explain altered bone matrix quality due to the administration of BPs in animal models INTRODUCTION: Non-enzymatic glycation (NEG) is a posttranslational modification of the organic matrix that results in the formation of advanced glycation end-products (AGEs). In bone, the accumulation of AGEs play an important role in determining fracture resistance, and elevated levels of AGEs have been shown to adversely affect the bone's propensity to brittle fracture. It was thus hypothesized that the suppression of tissue turnover in cortical bone due to the administration of bisphosphonates would cause increased accumulation of AGEs and result in a more brittle bone matrix. METHODS: Using a canine animal model (n = 12), we administered daily doses of a saline vehicle (VEH), alendronate (ALN 0.20, 1.00 mg/kg) or risedronate (RIS 0.10, 0.50 mg/kg). After a 1-year treatment, the mechanical properties, intracortical bone turnover, and the degree of nonenzymatic cross-linking of the organic matrix were measured from the tibial cortical bone tissue of these animals. RESULTS: There was a significant accumulation of AGEs at high treatment doses (+49 to + 86%; p < 0.001), but not at doses equivalent to those used for the treatment of postmenopausal osteoporosis, compared to vehicle. Likewise, postyield work-to-fracture of the tissue was significantly reduced at these high doses (-28% to -51%; p < 0.001) compared to VEH. AGE accumulation inversely correlated with postyield work-to-fracture (r (2) = 0.45; p < 0.001), suggesting that increased AGEs may contribute to a more brittle bone matrix. CONCLUSION: High doses of bisphosphonates result in the accumulation of AGEs and a reduction in energy absorption of cortical bone. The increased accumulation of AGEs in these tissues may help explain altered bone matrix quality due to the administration of BPs in animal models.

[Construction of mouse CCR3 gene RNAi lentivirus vector and its expression on mast cells].

Objective:The aim of this study is to screen the targeting chemokine receptor 3-RNA interference (CCR3-RNAi) lentiviral expression vector, infect mouse mast cells,observe the expression of this gene in mast cells and the interference efficiency of the virus vector.The pathogenesis of allergic rhinitis lays the foundation.Method:Three pairs of CCR3-shRNA sequences were constructed,and three pairs of double-stranded shRNA oligo were inserted into shRNA lentiviral vectors to construct three shRNA lentiviral recombinant plasmids.The recombinant vector and virus-packed auxiliary plasmids were co-transfected into 293T cells to obtain lentiviral plasmids.The lentiviral plasmids were then transfected into mouse bone marrow-derived mast cells in vitro and purified. The expression level of CCR3 mRNA in mast cells was verified by qRT-PCR,and the expression level of CCR3 protein in mast cells was detected by Western Blot.Result: It was confirmed by sequencing that the lentiviral vector of CCR3 shRNA was successfully constructed, transfected into 293T cells and packaged with virus. Finally the high purity PDSO19-PL-CCR3 lentiviral plasmid was obtained with a virus titer of 3.7×108TU/ml.The lentiviral plasmid was used to infect mouse mast cells.RT-PCR and Western Blot detection assay showed that CCR3shRNA reduced the expression of CCR3 gene in mouse mast cells at the level of mRNA and protein.Conclusion: The CCR3 gene RNAi lentivirus expression vector was successfully constructed.It was found that it downregulated the expression level of CCR3 gene mRNA and protein in mouse mast cells,which laid the foundation for further research on its role in the pathogenesis of allergic rhinitis.

L-carnitine protects against apoptosis of murine MC3T3-E1 osteoblastic cells.

L-carnitine (LC), an amino acid with a major role in cellular energy metabolism, has positive effects on bone metabolism. However, the effect of LC on apoptosis of osteoblast in vitro has not been reported. The aim of this study was to investigate the action of LC on apoptosis of mouse osteoblastic cell line MC3T3-E1. Cell apoptosis was measured by sandwich-enzyme-immunoassay. Release of cytochrome c from mitochondria into cytosol and Bcl-2, Bax protein levels were determined by Western blot analysis. The enzyme substrate was used to assess the activation of caspase-3 and caspase-9. LC inhibited MC3T3-E1 cell apoptosis induced by serum deprivation. Our study also shows that LC decreased cytochrome c release and caspase-3 and caspase-9 activation in serum-deprived MC3T3-E1 cells. Furthermore, LC protected against MC3T3-E1 cell apoptosis induced by the glucocorticoid (GC) dexamethasone (Dex).