Objective To investigate the cytidylyl phosphate guanosine(CpG) islands methylation status of E-cadherin (E-cad) promoter region in human ovarian carcinoma cell lines (ES-2,3 AO, SKOV3 ), and the effect of 5-azacytidine-2 '-deoxycytidines (5-Aza-CdR ) on the cell proliferative ability, invasion and the expression of E-cad protein. Methods Methylation specific PCR(MSP) was used to detect CpG islands methylation status of E-cad promoter region in ES-2,3AO and SKOV3 cell lines. After treated with different concentrations of 5-Aza-CdR, morphological changes of cell lines were observed under microscope. The proliferative ability was evaluated by methyl thiazolyl tetrazolium(MTT) assay. E-cad protein expression was detected by western-blot and cellular invasion was investigated by 24-well matrigel invasion chambers. Results Hypermethylatian status of CpG islands of E-cad promoter region was observed in ES-2 and SKOV3 cell lines, but not in 3AO cell lines. After treated with 5-Aza-CdR (0.1,1.0,10.0 μmol/L), ES-2 and SKOV3 cell lines displayed morphological evidence of differentiation. 5-Aza-CdR was found to decrease proliferation as evidenced by cell growth curve , to increase the level of E-cad protein expression (P ＜ 0.01 ), and effectively inhibit the ability of cell invasion(P ＜0.01 ). Conclusions CpG hypermethylation is an important mechanism of E-cad gene inactivation in ES-2 and SKOV3 cell lines. 5-Aza-CdR be found to inhibit proliferation and invasion, and increase the expression of E-cad probably by the inhibition of hypermethylation.

Carbon monoxide has been proved to be an important signal molecule in body. Transition metal carbonyl compounds are solidified form of carbon monoxide. Numerous studies have shown that Ruthenium carbonyl carbon monoxide releasing molecules have a strong pharmacological activity. In this paper, five Ruthenium (II) carbonyl CORMs 1-5 were synthesized and their toxicology, tissue distribution and interaction with blood endogenous substances were investigated. The results showed CORMs' IC50 to fibroblasts are ranged from 212.9 to 2089.2 micromol x L(-1). Their oral LD50 to mouse is between 800 to 1600 mg x kg(-1). After repeated administration, CORMs 1 and CORMs 5 haven't shown an obvious influence to rats' liver and kidney function, but caused the injury to liver and kidney cells. The in vivo distribution result revealed the majority of CORMs were distributed in blood, liver and kidney, only a small part of CORMs distributed in lung, heart and spleen. They could scarcely cross the blood-brain barrier and distribute to brain. The non-CO ligands in structure have an obvious relevance to their in vivo absorption and distribution. Interestingly, CORMs could enhance the fluorescence of bovine serum albumin, and this enhancement was in direct proportion with the concentration of CORMs. Under different conditions, interaction of CORMs with glutathione got different type of products, one is Ruthenium (II) tricarbonyl complexes, and Ruthenium (II) dicarbonyl complexes.

Objective To investigate the relationship between sleep fragmentation(SF)parameters and left ventricular structure in a community-based elderly population using data collected from the Multi-Ethnic Study of Atherosclerosis(MESA)cohort.Methods A total of 1 404 participants from the MESA cohort who underwent polysomnography(PSG)and cardiac magnetic resonance(CMR)were included.PSG was used to assess SF parameters,including wake after sleep onset(WASO),arousal index-total(ArI-total),arousal index-rapid eye movement(ArI-REM),and arousal index-non-rapid eye movement(ArI-NREM).Left ventricular end-diastolic mass(LVEDM)was measured via CMR.Linear regression was employed to analyze the relationship between SF parameters and left ventricular mass to height ratio(LVHi).Results Univariate linear regression analysis showed that WASO[β 0.134;95％confi-dence interval(CI)0.023-0.052;P＜0.001],ArI-total(β 0.184;95％CI 0.203-0.362;P＜0.001),ArI-REM(β 0.116;95％CI 0.100-0.260;P＜0.001)and ArI-NREM(β 0.175;9 5％CI 0.176-0.323;P＜0.001)were positively correlated with LVHi.After adjust-ment for confounding factors in multivariate linear regression analysis,WASO(β 0.045;95％CI 0.001-0.024;P=0.033)was positively correlated with LVHi.WASO(β 0.089;95％CI 0.006-0.034;P=0.006)was significantly associated with increased LVHi in the female group but not in the male group.Conclusion In an ethnically diverse cohort,WASO is significantly associated with increased LVHi after adjustment for potential confounders,especially in the female elderly population.

Objective: To evaluate the clinical effect of clear aligners in the anterior region in non-extraction cases by establishing a three-dimensional model of crown-root fusion to guide clinical application. Methods: Eleven patients visiting the orthodontic department of Xuzhou Stomatological Hospital from December 2020 to December 2021 were collected, and the orthodontic plan was designed using Maestro 3D Dental Studio scheduling software to obtain the expected three-dimensional model of the patient's orthodontic treatment result. CBCT, intraoral scan, and 3D reconstruction software were used to create a postoperative model of the patient. The crown and root data were aligned in Geomagic Studio 2014, and differences in torque and axial inclination between the actual model after treatment and the predicted model of the anterior teeth before treatment were compared in 3-matic. Results: The actual torque angles of the anterior teeth were all smaller than the predicted angles before treatment, with the highest realization rate of 77.55% for lateral incisors and the lowest of 60.70% for central incisors; the actual axial inclination angles of the anterior teeth were also smaller than the predicted angles before treatment, with the highest realization rate of 81.49% for central incisors and the lowest of 74.95% for cuspids. Conclusion A digital model of crown-root integration based on a combination of 3D reconstruction and intraoral scanning techniques is advantageous in assessing the efficacy of clear aligners. In non-extraction cases with clear aligner, the efficiency of movement is higher for small areas of the anterior region.

OBJECTIVETo study the expression of OCT4 protein in bladder cancer and its correlation to the clinicopathologic features and prognosis of bladder cancer.

METHODSOCT4 mRNA and protein expression was detected in 5 bladder cancer cell lines (RT-4, Tcc-Sup, KK47, T24, and 5637) and 1 normal bladder cell lines by real-time PCR and Western blotting, respectively. Immunohistochemical analysis was used to detect the expression of OCT4 protein in 46 bladder cancer samples.

RESULTSAll the 5 bladder cancer cell lines expressed detectable levels of OCT4 mRNA and proteins, whereas the normal bladder cell line SV-HUC-1 was negative for OCT4 expression. The clinical bladder cancer tissues showed a high positivity rate of OCT4 expression (76.1%), which was not detected in normal bladder tissues. Specific OCT-4 signals were localized mainly in the nuclei of the cancer cells. The expression rate of OCT4 protein was significantly higher in bladder cancer tissue than in normal bladder epithelium (P<0.05), and showed a positive correlation to the grade of tumor differentiation and metastasis (P<0.05) but not to the patients' age, gender or TNM stage.

CONCLUSIONOCT4 protein expression is associated with tumor differentiation and metastasis in bladder cancer and may play an important role in the early diagnosis and prognostic evaluation of bladder cancer.

Cell Line, Tumor ; Humans ; Neoplasm Metastasis ; Neoplasm Staging ; Octamer Transcription Factor-3 ; metabolism ; Prognosis ; Urinary Bladder Neoplasms ; diagnosis ; metabolism ; pathology

ObjectiveTo investigate the risk factors for failure in emergency endoscopic injection sclerotherapy (EIS) combined with sequential histoacryl injection (HI) for esophagogastric variceal bleeding (EGVB) with portal vein embolus (PVE). MethodsA total of 109 EGVB patients with PVE who underwent emergency gastroscopy in Beijing Shijitan Hospital, Capital Medical University, and The Fifth Medical Center of Chinese PLA General Hospital from January 2018 to December 2019 were enrolled, and according to the outcome of hemostatic treatment under emergency gastroscopy, the patients were divided into hemostatic failure group with 28 patients and hemostatic success group with 81 patients. The two groups were compared in terms of general information, varices and bleeding manifestations under gastroscopy, blood biochemical parameters, Child-Pugh class, and Model for End-Stage Liver Disease (MELD) score, and the risk factors for hemostatic failure were analyzed. The t-test or the Mann-Whitney U test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data between groups; a logistic regression analysis was used for multivariate analysis. ResultsCompared with the hemostatic success group, the hemostatic failure group had significantly higher peripheral white blood cell count (WBC), total bilirubin (TBil), Child-Pugh class, and MELD score (Z=3.794, Z=4.751, χ2=40.104, Z=6.412, all P＜0001) and significantly lower prothrombin time activity (PTA), albumin (Alb), and cholinesterase (CHE) (t=9.653, Z=3.093, Z=4.092, P＜0.001, P=0.002, and P＜0.001). WBC (odds ratio ［OR］=28.543, 95% confidence interval ［CI］: 1.285-634.113, P＜0.05), PTA(OR=0.194, 95%CI: 0.045-0.835, P＜0.05), TBil (OR=2.197, 95%CI: 1.004-4.810, P＜0.05), Alb (OR=0448, 95%CI: 0.209-0.961, P＜0.05), and Child-Pugh class (OR=5.164, 95%CI: 1.307-20.406, P＜0.05) were independent risk factors for hemostatic failure. ConclusionWBC, PTA, TBil, Alb, and Child-Pugh class are independent risk factors for failure in emergency EIS combined with sequential HI in the treatment of EGVB with PVE, and adequate preoperative evaluation and correction may help to improve the success rate of hemostasis.

Objective:To analyze, with Wuhan as an example, the problems found in the infrastructure development of China′s healthcare institutions, and put forward optimization suggestions for future epidemic prevention and control.Methods:From May to June 2020, we surveyed with questionnaires 56 healthcare institutions in Wuhan, covering such aspects as basic information of the institutions, infectious disease infrastructure readiness, and the converted wards for COVID-19. Statistical descriptions were used to analyze data so acquired.Results:The number of beds in the infectious departments of healthcare institutions in Wuhan amounted to 1.64 per 10 000 people, yet the existing 1 873 beds and about 5 000 convertible beds failed to meet the medical needs against the outbreak of COVID-19. After the outbreak, a total of 19 084 convertible beds were set up, of which general hospitals accounted for 88%; the area occupied by each converted bed in traditional Chinese medicine hospitals, hospitals of traditional and Western medicine, and maternal and child hospitals (<30.0m 2) was lower than that in general hospitals and specialist hospitals (>40.0m 2). Conclusions:Healthcare institutions should scientifically allocate " peacetime-wartime adaptive" hospital beds, optimize both the number and efficiency of these beds, and prepare for the worst scenarios, so that the infrastructure can be built and maintained in strict accordance with standards, government departments can rationally arrange infectious disease prevention and control facilities and strengthen their planning in case of emergencies.

Objective:To explore the feasibility in reconstruction of the muscular power with the superficial part of lateral femoral muscle through anatomical study on the superficial region of lateral femoral muscle.Methods:Studies on 4 sides of lower limbs of 2 cadaver specimen were conducted in the Department of Hand and Foot Microsurgery of Xi'an Fengcheng Hospital. Intraoperative observations and measurements were further carried out on 21 sides of 21 patients. Muscular fascia in superficial region, muscular gross morphology, thickness, length and width of muscles, length of muscle fibres and pinnate angles of muscle surface were observed and measured. Both blood vessels and nerves in the muscle were separated to measured.Results:The superficial region of lateral femoral muscle was in a shape of fusiform and started from the greater trochanter and ended at the patella and rectus femoris, with the fascia at proximal end and the muscle of distal end. The inferior muscle fibres of the fascia were arranged in sequence and ended at the deep fascia from proximal to distal. Mean muscle thickness was measured at 1.96 cm±0.48 cm, and mean pinnate angle was of 18.9°±3.3°. The superficial region was found being distributed by the descending branches of lateral circumflex femoral artery(LCFA) and the second branch of femoral nerve, and they accompanied each other. At 5.0 cm from the point of entry to the muscle, the diameter of the vessels was measured at 2.39 mm±0.52 mm, and the diameter of nerves was at 2.64 mm±0.61 mm. Both of arteries and nerves further branched out anteriorly and posteriorly in 1.0-1.5 cm intervals after having entered the muscle. At 0 - 2.5 cm away from the muscle entry point, a larger branch was often running posteriorly into the muscle, and this branch appears on all 4-sided specimens. While the occurrence rate in the 21 sides of patients observed in operations was of 90.5%, with a transverse diameter at 1.23 mm±0.28 mm.Conclusion:The superficial region of lateral femoral muscle is dominated by independent vessels and nerves and there are many branches from superior vessels and nerves, which form an anatomical basis for one or more muscular flaps.

Objective:To explore the effect of pentraxin 3 (PTX3) on memory improvement and Aβ expression in Alzheimer's disease (AD) model mice.Methods:(1) Ten 5-month-old 5×FAD mice were randomly divided into PTX3 group and model group ( n=5); 5 C57BL/6 wild-type mice at the same age were selected as control group; mice in the PTX3 group and control group were stereotactically injected 4 μL 0.5 g/L PTX3 or same dose of phosphate buffered saline (PBS); Morris water maze test was used to detect the learning and memory abilities, Y maze test was used to detect the short-term memory, and ELISA was used to obsevre the contents of Aβ 40 and Aβ 42 in the brain hemisphere. (2) Twenty-five 3-month-old 5×FAD mice were randomly divided into model group, 2 μg/kg PTX3 group, 4 μg/kg PTX3 group, 8 μg/kg PTX3 group, and 16 μg/kg PTX3 group ( n=5); 5 C57BL/6 wild-type mice at the same age were selected as control group; mice in the PTX3 groups were intranasally injected 2, 4, 8, and 16 μg/kg PTX3, respectively; those in the model group and control group were intranasally injected same dose of PBS; injection was given once every 96 h for a total of 7 times. Morris water maze test was used to detect the learning and memory abilities, Y maze test was used to detect the short-term memory, and ELISA was used to obsevre the contents of Aβ 40 and Aβ 42 in the hippocampus. Results:(1) Compared with the model group, the PTX3 group had significantly shorter platform latency, higher percentage of exploration time and higher percentage of spontaneous alternations ( P<0.05). Compared with those in model group ([63.38±21.42] pg/mL, [29.77±6.11] pg/mL), the concentrations of Aβ 40 and Aβ 42 in the brain tissues of PTX3 group ([15.87±2.11] pg/mL, [16.55±1.95] pg/mL) were statistically lower ( P<0.05). (2) Compared with the model group, the 16 μg/kg PTX3 group had significantly shorter escape latency and higher percentage of exploration time ( P<0.05); compared with the model group, the 2 μg/kg PTX3 group and 16 μg/kg PTX3 group had significantly higher percentage of spontaneous alternations ( P<0.05). The contents of Aβ 40 and Aβ 42 in the hippocampus of 8 μg/kg PTX3 group and 16 μg/kg PTX3 group were statistically lower compared with those in the model group ( P<0.05). Conclusion:PTX3 may attenuate cognitive deficits and decrease Aβ expression in the brain or hippocampus tissues of 5×FAD mice with AD.

BACKGROUND:Sema3A is a power secretory osteoprotective factor.However,studies about Sema3A-modified dental pulp stem cells(Sema3A-DPSCs)are rare. OBJECTIVE:To explore the osteogenic differentiation ability of Sema3A-DPSCs and their regulatory effect on the osteogenic differentiation of the pre-osteoblast cell line MC3T3-E1. METHODS:First,Sema3A-DPSCs were constructed using a lentivirus infection system carrying the Sema3A gene.Control lentivirus-treated DPSCs(Vector-DPSCs)were used as controls.Sema3A-DPSCs or Vector-DPSCs were co-cultured with proosteoblast line MC3T3-E1 at the ratio of 1∶1 and 1∶3 for 24 hours.Finally,the Sema3A-DPSCs,Vector-DPSCs and their co-cultured cells with MC3T3-E1 were cultured for osteogenic induction and differentiation.Osteogenic gene expression was detected by alkaline phosphatase staining,alizarin red staining and real-time quantitative RT-PCR to evaluate osteogenic differentiation ability. RESULTS AND CONCLUSION:(1)Sema3A mRNA and protein expression levels in Sema3A-DPSCs were significantly up-regulated.The level of secreted Sema3A in cell supernatant was up-regulated.(2)Compared with the Vector-DPSCs,mRNA expressions of osteogenic genes alkaline phosphatase,Runt-related transcription factor 2,osteocalcin and Sp7 transcription factors in Sema3A-DPSCs were up-regulated;the activity of alkaline phosphatase was enhanced,and the formation of mineralized nodules increased.(3)There were no obvious differences in proliferation between Sema3A-DPSCs and Vector-DPSCs.(4)Compared with MC3T3-E1/Vector-DPSCs co-culture system,the expression of MC3T3-E1 osteogenic genes was up-regulated,and the total alkaline phosphatase activity was enhanced and more mineralized nodules were formed in the MC3T3-E1/Sema3A-DPSCs co-culture system.(5)The results suggest that overexpression of Sema3A can enhance the osteogenic differentiation of DPSCs.Overexpression of Sema3A in DPSCs can promote osteogenic differentiation of MC3T3-E1 in the DPSCs/MC3T3-E1 co-culture system.