Objective To Study the effect of efflux pump-inhibitors(EPI)-Verapamil in photodynamic therapy (PDT) using hematoporphyrin monomethylether (HMME) as photosensitizer on the cariogenic bacteria in dental plaque biofilms.Methods According to the administrator order of the verapamil and photosensitizer in PDT,streptococcus mutans,streptococcus sanguis,eosinophilic lactobacillus and actinomyces viscosus were used to establish the dental plaque biofilm model.The experiment was divided into five groups,group A was incubated with the photosensitizer and verapamil group,group B using verapamil before incubated the photosensitize,group C suing photosensitizer before incubated verapamil,group D with PDT only,group E was control group.After laser treatment,the influence of the dental plaque biofilms was observed by confocal laser scanning microscope.Results As saline-treated group is a group of normal the dental plaque biofilms.In PDT only group,compared with the saline group,red fluorescence increased significantly,the bacteria lose accumulation capacity,and were isolated and scattered in dispersed state.In PDT plus verapamil group,compared with only PDT group,green staining increased,bacterial activity increased.In group B,cells were incubated with verapamil before incubated the photosensitizer group,green staining increased significantly,red fluorescence reduced,indicating live bacteria increased,and bacteria activity was significantly increased.Conclusion PDT is an effective method in eliminating cariogenic bacteria of dental plaque biofilms.Bacterial efflux pump inhibitors can lower HMME-PDT inhibition cariogenic bacteria in dental plaque biofilm,and pre-verapamil administration could significantly inhibit the effect of PDT treatment of dental caries.

Objective The purpose of this study was to assess the antibacterial effects of hematoporphyrin monomethylether-photodynamic therapy (HMME-PDT) on Enterococcus faecalis within infected simulated lateral canals in vitro with different energy.Methods Simulated lateral canals were prepared on extracted teeth.The specimens were infected with Enterococcus faecalis and then were randomly divided into eight groups.Group A was considered as negative control,its specimens were irrigated with physiological saline.Group B was the positive control,they were irrigated with 5.25％ NaClO.Other groups were incubated with HMME at concentration of 40 μg/ml for 5 min,followed by exposure to light at 532 nm for 120 s with different powers in a spiral pattern.Groups were named C-H,in corresponding with the power 50,60,70,80,90,100 mW.Microbial samples (the dentin chips from simulated lateral canals) were taken before and after the treatments.The survival fractions in each simulated lateral canal was calculated by counting colony-forming units(CFUs).Results 1.The HMME-PDT-treated groups resulted in a significant reduction in the number of E.faecalis in simulated lateral canals compared with the negative controls(P＜0.05).2.The antibacterial effects of group D、E、F、G、H were higher than the NaCl0-irrigated group(P＜0.05).3.The difference of antibacterial effects between the F、G、H groups was in significant(P＞0.05).Conclusion HMME-PDT had significant inhibitive efficacy on Enterococcus faecalis within infected simulated lateral canals and the efficacy was power-depended.It can play the best effect with the power of 80 mW.

Objective: To investigate the level of human blood basic fibroblast growth factor (FGF2) among children with autism spectrum disorder (ASD) and its correlation with behavioral phenotypes, to provide a reference for etiological research of ASD. Methods: ASD Children were selected to get rehabitation training in reseach center of children development behavior in Harbin Medical University and the rehabitation constitution for ASD disabilities in Heilongjiang, 40 children were induded as ASD group, 41 healthy children in Harbin kindergarten was classified as control group. The Autism Behavior Checklist (ABC), Childhood Autism Rating Scale (CARS) and Peabody Picture Vocabulary Test (PPVT) were used to assess the severity and intelligence of ASD children, respectively. Results: No difference was found in FGF2 level between ASD children (4.95 pg/mL) and normal children(8.51 pg/mL)（P>0.05). However, difference in FGF2 level between the two groups were found in 4-year-old group(P<0.05). The level of FGF2 differed across different severity and intelligence of ASD children(P<0.05). Conclusion Abnormal levels of FGF2 in ASD children may correlate with severity of autistic traits and intelligence of children.

Objective:To investigate the effect of lncRNA AC132217.4 on the proliferation and invasion of liver cancer MHCC97-H cells and its molecular mechanism.Methods:The TCGA database was used to analyze the differential expression of AC132217.4 in liver cancer tissue and adjacent tissue, and to analyze the relationship between the expression level of AC132217.4 and the overall survival of liver cancer patients. Transfection of pcDNA-AC132217.4 plasmid into MHCC97-H cells was defined as AC132217.4 group, transfection of pcDNA plasmid into MHCC97-H cells was defined as negative control (NC) group, respectively. The proliferation and invasion ability of MHCC97-H cells were detected by MTT method and Matrigel invasion assay. The binding site between AC132217.4 and miR-18a-5p was analyzed by starBase v2.0 software and dual luciferase reporter gene assay. Real-time quantitative PCR (RT-qPCR) detected the differential expression of miR-18a-5p in the two groups of MHCC97-H cells. The expression of epithelial-mesenchymal transition protein was detected by Western-blotting. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between the two groups. Results:Compared with adjacent tissues, the expression of AC132217.4 was down-regulated in liver cancer tissues ( P<0.01). Compared with liver cancer patients with low expression of AC132217.4, the overall survival of liver cancer patients with high AC132217.4 expression was longer ( P<0.05). The pcDNA-AC132217.4 plasmid significantly inhibited the proliferation of MHCC97-H cells ( P<0.05). The number of invasive cells in the NC group and AC132217.4 group were (131.30±12.55) and (37.45±7.77), respectively. The pcDNA-AC132217.4 plasmid significantly inhibited the invasive ability of MHCC97-H cells ( t=6.36, P<0.01). AC132217.4 directly complemented miR-18a-5p ( P<0.01). The expression of miR-18a-5p in MHCC97-H cells in AC132217.4 group (1.04±0.30) was significantly lower than that in NC group (6.13±0.75) ( t=6.27, P<0.01). Compared with the NC group, the expressions of epithelial phenotype proteins Cytokeratin and Claudin-1 in MHCC97-H cells in AC132217.4 group were up-regulated, while the expressions of mesenchymal phenotype proteins Vimentin, Slug and Snail were down-regulated. Conclusions:The expression of AC132217.4 is low in liver cancer tissue, and it is related to the overall survival of liver cancer patients. AC132217.4 might inhibit the proliferation and invasion of liver cancer MHCC97-H cells by sponge miR-18a-5p.

Objective To investigate the expression of miR-6768-5p in lung cancer tissue and its effect on the proliferation and invasion of lung cancer cells through targeted regulation of carboxypeptidase A4(CPA4).Methods The expression of miR-6768-5p in lung cancer tissues and adjacent tissues was analyzed u-sing the TCGA database.Quantitative real-time PCR(qPCR)was used to detect the expression of miR-6768-5p in human lung cancer cell lines(HCC1588,H1650,H1299,A549,HCC827)and normal alveolar epithelial cells(HPAEpiC cells).Lung cancer cells were transfected with NC mimics and miR-6768-5p mimics,respec-tively,and divided into NC group and miR-6768-5p group.The MTS assay and Matrigel invasion assay were used to detect the cell proliferation and invasion ability of each group,respectively.The putative binding sites of miR-6768-5p and CPA4 were verified using RNAhybrid software and dual-luciferase reporter gene experi-ment.The expression of CPA4 mRNA in each group of cells was detected by qPCR.The expression of AKT/c-MYC signaling pathway proteins in the cells of each group was analyzed by Western blot.Results Com-pared with the adjacent tissues,the relative expression level of miR-6768-5p in lung cancer tissues was signifi-cantly decreased,and the difference was statistically significant(P＜0.05).Compared with HPAEpiC cells,the relative expression level of miR-6768-5p was significantly decreased in lung cancer cell lines,and the differ-ence was statistically significant(P＜0.05).Compared with the NC group,the cell proliferation rate of miR-6768-5p group was significantly decreased(P＜0.05).The number of invasive cells in NC group and miR-6768-5p group was(131.30±12.55)and(37.45±7.77),respectively,and the number of invasive cells in miR-6768-5p group was significantly lower than that in NC group(P＜0.05).The relative expression level of CPA4 mRNA in H1299 cells of miR-6768-5p group was significantly lower than that in NC group(t=4.93,P＜0.05).Compared with the NC group,the expressions of AKT/c-myC signaling pathway proteins p-AKT,p-mTORC1,XIAP,MDM2 and C-myC proteins in miR-6768-5p group were significantly decreased.Conclusion The expression of miR-6768-5p is decreased in lung cancer tissues,and miR-6768-5p may inhibit the activation of AKT/c-MYC signaling pathway by targeting CPA4,and reduce the proliferation and invasion ability of lung cancer H1299 cells.

Objective: To identify the prevalence of sleep problems in children with autism spectrum disorder (ASD) and to explore the association with the main melatonin metabolite, 6-sulfatoxymelatonin (6-SM). Method: This was a prospective case-control study. Children with ASD were recruited from Child Development and Behavioral Research Center (CDBRC) of the Harbin Medical University and Harbin Special Education School from October 2015 to April 2017 (ASD group) . Healthy controls were selected from five kindergartens and one primary school in Harbin by the stratified cluster random sampling (control group) . The Children's Sleep Habits Questionnaire (CSHQ) was used to investigate the sleep problems of the two groups. The patients were matched in a 1∶1 ratio for the age and sex, and the urine samples of case-control pairs were collected in the morning. The level of 6-SM was measured by the enzyme linked immunosorbent assay (ELISA). The student's t test was used for comparison between the ASD group and control group, and the Pearson correlation analysis was used to determine the correlation difference. Result: A total of 212 ASD children (mean (±SD) age was (6.0±2.7) years, and 181 patients (85.4%) were male), and a total of 334 healthy children(mean (±SD) age was (5.9±2.6) years, and 272 patients (81.4%) were male) were recruited. Among them, 101 matched case-control pairs completed the collection of urine samples. According to the statistical analysis, the scores of total CSHQ, bedtime resistance, sleep onset delay, sleep duration, night waking, parasomnia, sleep disordered breathing and daytime sleepiness in children with ASD were significantly higher than those in the control group (48.2±6.2 vs. 46.6±5.4, 11.4±2.5 vs. 10.7±2.8, 1.7±0.8 vs. 1.5±0.7, 4.1±1.4 vs. 3.7±1.1, 4.2±1.5 vs. 3.8±1.1, 8.5±1.5 vs. 8.3±1.4, 3.7±1.0 vs. 3.4±0.8, 11.7±2.5 vs. 12.4±2.7, t=3.16, 3.00, 3.23, 2.76, 3.19, 1.99, 3.45,-2.72, P=0.002, 0.003, 0.001, 0.006, 0.002, 0.048, 0.001, 0.007), the level of 6-SM was significantly lower in children with ASD than that of healthy controls ((1.24±0.50) vs. (1.68±0.63)μg/h, t=-5.50, P<0.01), and the total CSHQ score was negatively correlated with the level of 6-SM (r=-0.50, P<0.01). Conclusion The children with ASD were at high risk for sleep problems, and the melatonin metabolite of ASD group was abnormal compared with that of the control group. Moreover, there was a negative correlation between the severity of sleep problems and the level of 6-SM in ASD children. The results of our study indicate that the abnormal melatonin metabolism may be one of the causes of sleep problems in children with ASD.

Objective To construct the Markov models to reflect the reality of prevention and treatment interventions against hepatitis B virus (HBV) infection,simulate the natural history of HBV infection in different age groups and provide evidence for the economics evaluations of hepatitis B vaccination and population-based antiviral treatment in China.Methods According to the theory and techniques of Markov chain,the Markov models of Chinese HBV epidemic were developed based on the national data and related literature both at home and abroad,including the settings of Markov model states,allowable transitions and initial and transition probabilities.The model construction,operation and verification were conducted by using software TreeAge Pro 2015.Results Several types of Markov models were constructed to describe the disease progression of HBV infection in neonatal period,perinatal period or adulthood,the progression of chronic hepatitis B after antiviral therapy,hepatitis B prevention and control in adults,chronic hepatitis B antiviral treatment and the natural progression of chronic hepatitis B in general population.The model for the newborn was fundamental which included ten states,i.e.susceptiblity to HBV,HBsAg clearance,immune tolerance,immune clearance,low replication,HBeAg negative CHB,compensated cirrhosis,decompensated cirrhosis,hepatocellular carcinoma (HCC) and death.The susceptible state to HBV was excluded in the perinatal period model,and the immune tolerance state was excluded in the adulthood model.The model for general population only included two states,survive and death.Among the 5 types of models,there were 9 initial states assigned with initial probabilities,and 27 states for transition probabilities.The results of model verifications showed that the probability curves were basically consistent with the situation of HBV epidemic in China.Conclusion The Markov models developed can be used in economics evaluation of hepatitis B vaccination and treatment for the elimination of HBV infection in China though the structures and parameters in the model have uncertainty with dynamic natures.

Objective:To construct an evaluation index system for the risk factors of peripheral venous indwelling catheter failure in adults.Methods:Through literature review, focus group interview method and Delphi method, an evaluation index system for risk factors of peripheral venous indwelling catheter failure in adults was established.Results:The recovery rate of the two round were both 100%; the authority coefficient was 0.939, the coordination coefficient was 0.452 and 0.315 respectively ( χ2=607.88, 502.45, P<0.05). Finally an evaluation index system for adult peripheral venous indwelling needle catheter failure risk factors including puncture factors, patient factors, indwelling needle placement factors, indwelling needle maitenance and withdrawal factors and 54 second-level indicators were formed. Conclusions:The construction of adult peripheral venous indwelling needle catheter failure risk factor assessment system is reasonable and comprehensive in content, which provides reference for clinical nurses to evaluate and prevent the failure of adult peripheral venous catheter failure.

Objective:To investigate the incidence rate and gene variation of methylmalonic academia (MMA) in Ji′nan city by analyzing biochemical and genetic screening results, and to explore the carrier frequency of MMA-related pathogenic genes in the population in Ji′nan.Methods:The children diagnosed with MMA by tandem mass spectrometry screening in Ji′nan Neonatal Disease Screening Centre from May 2011 to May 2022 were enrolled in this study.Their genetic test results were retrospectively analyzed and summarized.The dried heel blood tablets collected from 6 800 newborns were tested for neonatal gene screening. MMAA, MMAB, MMACHC and MMUT genes in 4 800 cases were detected by high-throughput sequencing+ target area capture technology.Ultra-multiplex polymerase chain reaction+ target gene locus capture technology was used to detect 174 target loci of 8 genes related to MMA in 2 000 cases.The hotspot mutation and related gene carrier rate of MMA were analyzed. Results:A total of 367 452 newborns were screened by tandem mass spectrometry, and 103 cases (56 males and 47 females) were diagnosed with MMA by screening.The estimated incidence of MMA was 1∶3 567.Among the 103 MMA cases, 76 were genetically diagnosed, and 4 gene variants of MMA ( MMAHC, MMUT, MMAA, MMADHC) were identified.A total of 6 800 neonates underwent neonatal genetic screening.Three of them were diagnosed with MMA.About 318 infants carried pathogenic variants of MMA, with a total carrier rate of 4.68%.Specifically, the carrier rates of MMACHC and MMUT gene variants were 3.09%(210/6 800) and 1.43% (97/6 800), respectively. Conclusions:MMA is the most common organic acid metabolism disorder in our country.The incidence and carrier rate of this disease are high in Jinan city.Neonatal genetic screening is an important supplement to neonatal biochemical screening.Carrier screening for MMA-related pathogenic genes is recommended for couples of childbearing age in Jinan.

Objective:To analyze the risk factors for postoperative one-year mortality in elderly patients with femoral intertrochanteric fracture following multidisciplinary treatment (MDT) by intramedullary nailing.Methods:The clinical data were retrospectively analyzed of the 158 elderly patients with femoral intertrochanteric fracture who had undergone MDT by proximal femoral intramedullary nailing between January 2018 and August 2020 at Department of Orthopedics, Trauma Center, Zhongda Hospital Affiliated to Southeast University. There were 41 males and 117 females with an average age of 82.5 years (from 65 to 95 years). By the modified Evans classification, there were 15 cases of type Ⅰ, 16 cases of type Ⅱ, 35 cases of type Ⅲ, 81 cases of type Ⅳ, and 11 cases of type Ⅴ. The one-year mortality was documented in the patients after surgery. To screen for risk factors, univariate analysis was conducted of gender, age, body mass index (BMI), modified Evans classification of fractures, time from injury to operation, American Society of Anesthesiologists (ASA) classification, Charlson comorbidity index (CCI) and comorbidities, as well as preoperative hemoglobin (Hb), serum albumin (ALB) and total lymphocyte count (TLC). The factors with P<0.05 were included in the multivariate logistic regression model analysis to determine the risk factors. Results:A total of 13 patients died within one year after surgery, yielding a mortality of 8.2% (13/158). Univariate analysis showed significant differences in age, body mass index, modified Evans classification of fractures, CCI and Hb between the surviving and dead patients ( P<0.05). Multivariate logistic regression analysis showed that age >85 years ( OR=0.122, 95% CI: 0.018 to 0.834, P=0.032), BMI>23.9 kg/m 2 ( OR=0.083, 95% CI: 0.013 to 0.510, P=0.007), CCI≥3 points ( OR=0.051, 95% CI: 0.090 to 0.275, P=0.001) and preoperative Hb<90 g/L ( OR=4.733, 95% CI: 1.036 to 21.624, P=0.045) were the independent risk factors for postoperative one-year mortality in the elderly patients with intertrochanteric fracture following MDT by proximal femoral intramedullary nailing. Conclusions:After MDT by proximal femoral intramedullary nailing of femoral intertrochanteric fractures, the geriatric patients with an age >85 years, BMI>23.9 kg/m 2, CCI≥3 points and Hb<90 g/L are likely to die. Therefore, special care should be taken for them.