BACKGROUND: Knee osteoarthritis (OA) is still challenging to prevent or treat. Enhanced endoplasmic reticulum (ER) stress and increased pyroptosis in chondrocytes may be responsible for cartilage degeneration. This study aims to investigate the effect of ER stress on chondrocyte pyroptosis and the upstream regulatory mechanisms, which have rarely been reported. METHODS: The expression of the histone methyltransferase enhancer of zeste homolog 2 (EZH2), microRNA-142-3p (miR-142-3p), and high mobility group box 1 (HMGB1) and the levels of ER stress, pyroptosis, and metabolic markers in normal and OA chondrocytes were investigated by western blotting, quantitative polymerase chain reaction, immunohistochemistry, fluorescence in situ hybridization, fluorescein amidite-tyrosine-valine-alanine-aspartic acid-fluoromethyl ketone (FAM-YVAD-FMK)/Hoechst 33342/propidium iodide (PI) staining, lactate dehydrogenase (LDH) release assays, and cell viability assessments. The effects of EZH2, miR-142-3p, and HMGB1 on ER stress and pyroptosis and the hierarchical regulatory relationship between them were analyzed by chromatin immunoprecipitation, luciferase reporters, gain/loss-of-function assays, and rescue assays in interleukin (IL)-1β-induced OA chondrocytes. The mechanistic contribution of EZH2, miR-142-3p, and HMGB1 to chondrocyte ER stress and pyroptosis and therapeutic prospects were validated radiologically, histologically, and immunohistochemically in surgically induced OA rats. RESULTS: Increased EZH2 and HMGB1, decreased miR-142-3p, enhanced ER stress, and activated pyroptosis in chondrocytes were associated with OA occurrence and progression. EZH2 and HMGB1 exacerbated and miR-142-3p alleviated ER stress and pyroptosis in OA chondrocytes. EZH2 transcriptionally silenced miR-142-3p via H3K27 trimethylation, and miR-142-3p posttranscriptionally silenced HMGB1 by targeting the 3'-UTR of the HMGB1 gene. Moreover, ER stress mediated the effects of EZH2, miR-142-3p, and HMGB1 on chondrocyte pyroptosis. In vivo experiments mechanistically validated the hierarchical regulatory relationship between EZH2, miR-142-3p, and HMGB1 and their effects on chondrocyte ER stress and pyroptosis. CONCLUSIONS A novel EZH2/miR-142-3p/HMGB1 axis mediates chondrocyte pyroptosis and cartilage degeneration by regulating ER stress in OA, contributing novel mechanistic insights into OA pathogenesis and providing potential targets for future therapeutic research.
Enhancer of Zeste Homolog 2 Protein/genetics* ; Osteoarthritis, Knee/pathology* ; Chondrocytes/metabolism* ; Pyroptosis/physiology* ; HMGB1 Protein/genetics* ; MicroRNAs/metabolism* ; Endoplasmic Reticulum Stress/genetics* ; Humans ; Animals ; Rats ; Male ; Rats, Sprague-Dawley ; Middle Aged

ObjectiveAn analytical method was developed for the qualitative screening of 52 illegally adulterated weight-losing compounds in foods by ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. MethodsThe samples were extracted by methanol. Waters CORTECS T3 column （100 mm×2.1 mm， 2.7 μm） was used with acetonitrile -0.1% formic acid aqueous solution as mobile phases by gradient elution. The compounds were detected by electrospray ionization mass spectrometry and Q-TOF-MS detector in positive and negative ion mode. A standard spectrum library was established by reference standards， and the qualitative analysis was finished by the comparison of the retention time， parent ion and fragment ion accurate molecular mass of each compound in the sample and the library. ResultsThe method was specific without interference of blank matrix， and repeatable in sextuplicate. The detection limits of 52 compounds in 5 matrix were 1‒100 mg‧kg^-1. The method was successfully applied to the analysis of actual samples，and 16 compounds were checked out in 246 samples. ConclusionThe method is accurate， specific and sensitive， which can be used to combat the illegal adulteration behavior effectively.

Objective: To establish a hypertension risk assessment model among the middle-aged and elderly populations based on residents' electronic healthcare records of the basic public health service program, so as to provide insights into prevention of hypertension. Methods: Demographic features and physical examinations were collected among residents at ages of 40 years and older from residents' electronic healthcare records of the basic public health service program in a county of Zhejiang Province from 2019 to 2020. The risk factors of hypertension were identified using a multivariable logistic regression model, and the odds ratio (OR) for each risk factor was transformed into approximate relative risk (RR), which was included in the formula for calculation of the disease risk proposed by Harvard School of Public Health to create a hypertension risk assessment model. The predictive value of the model was evaluated using a receiver operator characteristic (ROC) curve. Results: Totally 7 275 subjects were enrolled, with a mean age of (66.15±7.91) years, and the participants included 3 189 males and 4 086 females, with a male-to-female ratio of 0.78∶1. There were 190 cases with new-onset hypertension (2.61%). Multivariable logistic regression analysis revealed that overweight, obesity, central obesity, borderline high triacylglycerol (TG), elevated TG, abnormal fasting plasma glucose (FPG), prehypertension and family history of hypertension were included in the hypertension risk assessment model, with approximate RR values of 1.66, 1.96, 1.54, 1.17, 1.64, 1.45, 1.69 and 1.11. The area under the ROC curve (AUC) of the model was 0.678 (95%CI: 0.641-0.715, P<0.001), and the optimal positive cut-off was 0.899. The model predicted 139 subjects with RR>0.899 for hypertension, with a sensitivity of 73.16% and specificity of 55.79%. Conclusions The hypertension risk assessment model created in this study is feasible to predict the RR for developing hypertension among the middle-aged and elderly populations, which has a predictive value in healthcare management.

OBJECTIVE: To analyze the clinical characteristics and genetic basis of a child affected with Glass syndrome. METHODS: Clinical manifestations and auxiliary examination results of the child were analyzed. Potential mutation was detected with next generation sequencing and validated by Sanger sequencing. RESULTS: The child has featured growth and mental retardation, delayed speech, cleft palate, crowding of teeth, and downslanting palpebral fissures. DNA sequencing revealed a de novo heterozygous missense mutation c.1166G>A (p.R389H) in exon 8 of the SATB2 gene in the child. CONCLUSION The heterozygous mutation c.1166G>A (p.R389H) of the SATB2 gene probably account for the Glass syndrome in the patient.
Abnormalities, Multiple ; genetics ; Child ; Chromosome Deletion ; Chromosomes, Human, Pair 2 ; Humans ; Intellectual Disability ; genetics ; Matrix Attachment Region Binding Proteins ; genetics ; Mutation ; Transcription Factors ; genetics

Objective: To investigate and analyze the awareness, utilization, and satisfaction of patients with chronic diseases (hypertension and type 2 diabetes mellitus), as well as the influencing factors related to health management services for chronic diseases in basic public health service projects in Zhejiang Province, in order to promote the development of effective health management services for chronic diseases. Methods: Between September and November 2017, 960 local patients with chronic diseases aged 35 years or older were randomly selected in eight counties (cities/districts) in Zhejiang province using the random number table, based on the chronic disease information from the information management system. A total of 394 (41.04%) male and 566 (58.96%) female patients, with an average age of (68.02±10.02) years, participated in this study. Face-to-face questionnaires were used to collect patients’ awareness, utilization, and satisfaction with health management services for chronic diseases. The awareness of health management services for chronic diseases and their utilization status were described and analyzed by statistical rate indicators. Possible factors affecting the awareness and utilization status were analyzed by χ² test and logistic regression. Satisfaction scores of patients with chronic diseases were analyzed by means and standard deviation. T-test was used to compare the satisfaction level with service items (service convenience, service environment, waiting time, medical skill level, and service attitude) between urban and rural patients with chronic diseases. Finally, multivariate logistic regression analysis was conducted to determine overall satisfaction. Results: The overall awareness of health management services for chronic diseases in chronic disease patients was 96.15%. Multivariate Logistic regression analysis showed that patients who lived in developed regions, rural areas, and patients who did not live alone had more awareness of health management services (OR= 0.347, 3.563, and 0.281, all P <0.05). The overall utilization rate of health management services for chronic diseases in chronic disease patients was 94.58%. Multivariate Logistic regression analysis revealed that patients who lived in developed regions and rural areas showed better utilization of health management services (OR= 0.297 and 4.145, all P<0.05). The average satisfaction score of patients for all items of health management services was above nine. The satisfaction scores of chronic disease patients in rural areas regarding service convenience, service environment, waiting time, and service attitude were better than those of chronic disease patients in urban areas (t=-2.157, -3.614, -3.164, -2.879, all P<0.05). The overall satisfaction score regarding health management services for chronic diseases was 92.95%. Multivariate Logistic regression analysis showed that patients who purchased health insurance for urban and rural residents reported higher overall satisfaction with health management services for chronic diseases compared to those patients who purchase other medical insurance (OR=2.943, P<0.05). Conclusion The current level of awareness, utilization, and satisfaction with health management services for chronic diseases among chronic disease patients in Zhejiang province should be further improved. Moreover, attention should be paid to the balanced development of urban and rural areas to further increase the equality level of basic public health services.

Objective To explore the financial compensation strategy on the basis of understanding the current situation of compensation for public health services at public hospitals in Zhejiang province.Methods From July to October,2017,an interview survey was conducted at 23 public hospitals in 7 cities in Zhejiang province to understand the current situation of financial compensation for public health services at public hospitals.At the same time,the data of income and expenditure in 2016 of every hospital were collected and analyzed by the methods of descriptive statistics.Results The average proportion of financial subsidies at the 23 public hospitals in 2016 was 6.29％,of which public health special subsidy accounted for 0.44％ of total hospital revenues.Only 9 of the 23 hospitals had surplus in 2016,accounting for 39.13％.Conclusions The public health services compensation mechanism of public hospitals in Zhejiang province calls for further reforms.The government departments should comprehensively formulate financial compensation policies at the provincial level according to the public health assignments and call into play a variety of compensation methods,to establish a long-term assessment mechanism and ensure that public hospitals can proactively carry out public health services.

Objective To evaluate the carotid and left ventricular function changes in patients with subclinical atherosclerosis (SA) using echo-tracking (ET) and three-dimensional speckle tracking echocardiography (3D-STE).Methods Eighty patients with SA were divided into low-risk group (27 cases),middle-risk group (26 cases) and high-risk group (27 cases) according to the Framingham risk score (FRS).Each of them was examined by echocardiography and carotid ultrasound to obtain the parameters including carotid intima-media thickness (cIMT),stiffness parameter (β),Pressure-strain elastic modulus (Ep),pulse wave conducting velocity(PWVβ),LV peak systolic global longitudinal strain (GLS),LV peak systolic global circumferential strain (GCS),myocardial wall stress (MWS) and left ventricular ejection fraction(LVEF) for analysis.The study got approval from the Ethics Committee of Tongji Medical College,Huazhong University of Science and Technology (NO:IORG00371).Results Ep,left atrial volume (LAV) and MWS had significantly differences among the three groups (all P ＜0.05).Compared with low-risk group,cIMT,β,PWVβ were higher and GLS was lower in high-risk group (all P ＜0.05).There were no statistical difference in LVEF and GCS among the three groups (all P ＞ 0.05).β was positively correlated with age and FRS,and negatively correlated with GLS (all P ＜ 0.05).LAV was positively correlated with age and E/e (all P ＜0.05).GLS was negatively correlated with FRS and β (all P ＜ 0.05).MWS was positively correlated with β and SBP,and negatively correlated with LVEF (all P ＜0.01).Conclusions ET combined with 3D-STE could be applied to evaluate the carotid and left ventricular function accurately in patients with subclinical atherosclerosis,and provide scientific bases for establishing intervention strategy.

This study aimed to examine the preparation of cationic lipid microbubble (CLM), and evaluate its physical and chemical properties and toxicity, measure the gene transfection efficiency by ultrasound triggered microbobble destruction (UTMD) in combination with CLM. The CLM was prepared by the method of the thin film hydration, and its morphology was observed under the electron microscopy at 1st, 3rd, 7th, 10th, and 14th day after preparation, respectively. The size, Zeta potential and stability of CLM were tested. The acute toxicity of CLM was assessed. The green fluorescent protein gene (EGFP) transfection efficiency was evaluated. The experiment grouping was as follows: naked plasmid group (P group), ultrasonic irradiation plus naked plasmid group (P-US group), naked plasmid plus CLM group (P-CLM group), naked plasmid plus ultrasound and CLM group (UTMD group). The expression of EGFP was detected by fluorescent microscopy and flow cytometry. The results showed that CLMs were spherical in shape, with the similar size and good distribution degree under the light and electron microscopies. The size of CLMs was varied from 250.4±88.3 to 399.0±99.8 nm and the Zeta potential of CLMs from 18.80±4.97 to 20.1±3.1 mV. The EGFP expression was the strongest in the UTMD group, followed by the P-CLM group, P-US group and P group. Flow cytometry results were consistent with those of fluorescent microscopy. The transfection efficiency was substantially increased in the P-US group, P-CLM group and UTMD group as compared with that in the P group, almost 7 times, 10 times and 30 times higher than that in the P group respectively. It is suggested that CLMs prepared by the method of thin film hydration are uniform in diameter, and proved non-toxic. UTMD combined with CLM can significantly increase the transfection efficiency of EGFP to targeted cells.

This study aimed to examine the preparation of cationic lipid microbubble (CLM),and evaluate its physical and chemical properties and toxicity,measure the gene transfection efficiency by ultrasound triggered microbobble destruction (UTMD) in combination with CLM.The CLM was prepared by the method of the thin film hydration,and its morphology was observed under the electron microscopy at 1st,3rd,7th,10th,and 14th day after preparation,respectively.The size,Zeta potential and stability of CLM were tested.The acute toxicity of CLM was assessed.The green fluorescent protein gene (EGFP) transfection efficiency was evaluated.The experiment grouping was as follows:naked plasmid group (P group),ultrasonic irradiation plus naked plasmid group (P-US group),naked plasmid plus CLM group (P-CLM group),naked plasmid plus ultrasound and CLM group (UTMD group).The expression of EGFP was detected by fluorescent microscopy and flow cytometry.The results showed that CLMs were spherical in shape,with the similar size and good distribution degree under the light and electron microscopies.The size of CLMs was varied from 250.4±88.3 to 399.0±99.8 nm and the Zeta potential of CLMs from 18.80±4.97 to 20.1 ±3.1 mV.The EGFP expression was the strongest in the UTMD group,followed by the P-CLM group,P-US group and P group.Flow eytometry results were consistent with those of fluorescent microscopy.The transfection efficiency was substantially increased in the P-US group,P-CLM group and UTMD group as compared with that in the P group,almost 7 times,10 times and 30 times higher than that in the P group respectively.It is suggested that CLMs prepared by the method of thin film hydration are uniform in diameter,and proved non-toxic.UTMD combined with CLM can significantly increase the transfection efficiency of EGFP to targeted cells.

BACKGROUND: Traditional glass ion sealant has a poor abradability and a low rupture strength. The sealant on the occlusal surfaces easily fell off, and is difficult to replace resin sealant. OBJECTIVE: To observe the effects of traditional resin sealant and atraumatic restorative treatment (ART) glass ionomer-based pit and fissure sealant for the young children. METHODS: Randomized comparison method was used to compare ART glass ionomer-based pit on molars of one side with resin sealant on the opposite side in 89 3-year-old children. RESULTS AND CONCLUSION: The retention rates of ART glass ionomer sealant after 6 and 18 months were significantly lower than those of resin sealant (P < 0.05). The caducous position of ART gliass ionomer sealant was the second deciduous molar of the lower mandible, but the caducous position of resin sealant was the second deciduous molar of the upper mandible. The secondary caries rate of ART glass ionomer sealant was significantly lower than that of resin sealant at 6 months. No significant difference was determined between groups at 18 months. These suggest that ART glass ionomer pit and fissure sealant has lower drop-out rate, simple operation and low cost with excellent caries-preventing effect. Since it is economically superior to resin sealant, the method is worth popularizing in caries-preventing projects.