Objective To construct a eukaryotic expression plasmid carrying the PKCI-1/HINT1 gene,to investigate its expression in A375 melanoma cells after transfection,and to evaluate its effects on apoptosis and autophagy of A375 cells.Methods The PKCI-1/HINT1 gene sequence was amplified by reverse transcription PCR (RT-PCR) with total RNA extracted from A375 cells as the template,then inserted into the eukaryotic expression plasmid PCDNA3.1 (+) to construct a recombinant plasmid,PCDNA3.1 (+)-PKCI-1/HINT1.Some A375 cells were classified into two groups to be transiently transfected with the recombinant plasmid (PCDNA3.1 (+)-PKCI-1/HINT1 group) or the empty plasmid PCDNA3.1 (+) (control group).After additional 48-hour culture,RT-PCR and Western blot analysis were performed to quantify the mRNA and protein expressions of PKCI-1/HINT1 respectively,Hoechst 33342 staining was conducted to detect apoptosis of A375 cells,Western blot analysis to detect the expressions of intracellular caspase-3 and autophagy-associated protein beclin1,and cell autophagy was observed by using the green fluorescent protein (GFP)-microtubule-associated protein 1 light chain 3 (LC3) labelling method combined with confocal laser scanning microscopy.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of A375 cells at 24,48,72 and 96 hours after transfection.Results Enzyme digestion and sequencing analysis confirmed that the eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed and effectively expressed in the transfected A375 cells.MTT assay showed that PKCI-1/HINT1 could obviously inhibit the proliferation of A375 cells,and the number of live cells was decreased by 17.0％,25.6％ and 29.4％ in the PCDNA3.1 (+)-PKCI-1/HINT1 group at 48,72 and 96 hours,respectively,compared with the control group (all P ＜ 0.05).Hoechest 33258 staining revealed that PKCI-1/HINT1 could promote the formation of apoptotic bodies in A375 cells.Confocal laser scanning microscopy demonstrated that the overexpression of PKCI-1/HINT1 increased GFP-LC3 puncta formation in A375 cells.In addition,Western blot analysis indicated that PKCI-1/HINT1 up-regulated the protein expressions of caspase-3 and beelin1 in A375 cells.Conclusions The eukaryotic expression plasmid PCDNA3.1 (+)-PKCI-1/HINT1 was successfully constructed,and PKCI-1/HINT1 could be effectively expressed in A375 cells.High-level expression of PKCI-1/HINT1 could suppress cellular proliferation,promote apoptosis,and induce autophagy,of A375 cells.

Objective To explore the characteristics of illness attribution of outpatients with multiple somatic symptoms in Peking Union Medical College Hospital. Methods It was a cross-sectional study conducted from March to October,2012. A total of 150 outpatients were recruited from the departments of Gastroenterology,Traditional Chinese Medicine and Psychological Medicine by convenience sampling. Somatic symptom scale of the Patient Health Questionnaire (PHQ-15) was used to screening each patient in the waiting list. With the cut-off value of 10,patients were divided into the somatic symptom positive (SOM+) group and somatic symptom negative (SOM-) group. Sociodemographic characteristics were compared between these two groups. All the subjects completed interviews including questions about illness attribution. All the answers of illness attribution were concluded into three major groups as physical factors,situational factors and psychological factors. Results The proportion of female was significantly higher in SOM+ group than in SOM-group (69.3% vs. 53.3%;χ=4.048,P=0.044). In SOM+ group,significantly more patients contributed their illness to psychological factors (64.0% vs. 45.0%;χ=5.273,P=0.022). There was no significantly difference between SOM+ group and SOM-group on the phenomenon of multiple illness attribution (41.0% vs. 32.0%;χ=1.407,P=0.236). However,in the Department of Gastroenterology,there were significantly more outpatients in SOM+ group with multiple illness attribution (60.0% vs. 32.0%;χ=3.945,P=0.047).Conclusions The outpatients in general hospital with multiple somatic symptoms are more likely to contribute their illness to psychological factors. The phenomenon of multiple illness attribution is common among patients. Clinicians should increase their awareness and knowledge of illness attribution,so as to provide better holistic health services.

Objective To investigate the MRI findings of perihip heterotopic ossification (HO) in the early, mid and late stages. Meth-ods The MRI of 44 inpatients with HO from February, 2011 to September, 2013 were reviewed, in which 20 cases (28 joints) were in early stage, 18 cases (24 joints) in mid stage and 6 cases (8 joints) in late stage. For the enhanced T1WI, 9 cases (11 joints) were in early stage, 6 cases (7 joints) in mid stage, and 3 cases (4 joints) in late stage. Theχ2 trend test was used to evaluate the MRI signal change with the HO maturity. Results With the maturity of hip HO, the signal intensity of T2WI reduced (χ2=16.773, P<0.001), fat signal on T1WI increased, the enhancement reduced (χ2=16.048, P=0.007). Conclusion The MRI findings of perihip HO are characteristic in MRI in all the stages. MRI is useful for the diagnosis of perihip HO, especially for the early HO.

AIM:To investigate the effect of quercetin on the biological functions of rat bone marrow-derived endothelial progenitor cells (EPCs) and its potential mechanisms.METHODS:The bone marrow-derived mononuclear cells of Sprague-Dawley rats were isolated by density gradient centrifugation.The differentiated EPCs were cultured specially and stained with DiI-Ac-LDL and FITC-UEA-1.CD133+ and FLK-1+ were detected on the cell surfaces.After 14 d, the EPCs were incubated with a PI3K inhibitor BYL719 (3 μmol/L) and an ERK inhibitor FR180204 (15 μmol/L).After incubation of the inhibitors for 2 h, the cells were treated with quercetin at different concentrations (0, 10, 20, 40, 80 and 100 μmol/L).MTT assay and Transwell assay were used to detect cell viability and the number of migratory cells.The protein levels of AKT, eNOS, ERK and their phosphorylated status were determined by Western blot.RESULTS:Quercetin enhanced the viability and migration of the EPCs at a dose-dependent manner.However, the PI3K inhibitor BYL719 suppressed the QUE-induced cell viability and migration.Moreover, ERK inhibitor FR180204 exerted the similar inhibitory effect on the cell viability but had no effect on cell migration.Quercetin activated the phosphorylation of AKT, eNOS and ERK.On the other hand, BYL719 was observed to inhibit the phosphorylation of AKT and ERK.FR180204, however, was showed to inhibit the phosphorylation of ERK only.On the contrast, the stimulatory effects that quercetin exerted on the expression of eNOS and its phosphorylation were suppressed by BYL719 and FR180204.CONCLUSION:Quercetin stimulates the viability and migration of EPCs via PI3K/AKT/eNOS and ERK/eNOS signaling pathway, which would be beneficial for cardiovascular health.

ObjectiveTo investigate the imaging findings of Charcot neuroarthropathy. Methods9 cases with Charcot neuroarthropathy determined clinically or pathologically were reviewed. ResultsThe findings of X-ray and CT including: soft tissue swelling in 9 cases, bone absorption in 5, bone proliferation in 7, periosteal reaction in 7, ectopic calcification or ossification in 9, luxation or semiluxation in 4. For MRI: soft tissue swelling, edema of joint capsule in 2, bone marrow dropsy in 1, sclerotin damage in 1. The joint capsule and periarticular soft tissue unevenly enhanced after Gd-DTPA. Nuclear medicine displayed radioactivity aggregation in 1 case. ConclusionX-rays plain film is the first choice for the diagnosis of Charcot neuroarthropathy, while CT, MRI and nuclear medicine may be helpful.

Objective: To examine the influencing factors of the efficiency of county-level centers for disease control and prevention ( CDCs) in China. Methods:458 county-level CDCs were selected based on a systematic sam-pling method. Multilevel modeling was used to analyze the region-level and institution-level influencing factors affect-ing the efficiency of CDCs. Results: It was found that the region ( province) is associated with the efficiency of a CDC. The region-level factor of population density exhibited a significant influence, while the institution-level factors such as the proportion of health technicians, service income and CDC laboratories per capita also had an influence on overall efficiency. Conclusions: Both the region-level and institution-level determinants influence efficiency. Multi-level modeling can help researchers gain a comprehensive understanding of the influencing factors that affect the CDC efficiency.

AIM: To investigate the role of transcription factor hairy and enhancer of split 1 (Hes1) in the malignant transformation of human bronchial epithelial cell line BEP2D induced by tobacco.METHODS: The BEP2D cells were chronically exposed to cigarette smoke condensate (CSC) at 1 cigarette per L until the 70th generation.The phenotype of malignant transformation of the cells induced by CSC was detected by soft agar clony formation assay.RT-PCR and Western blot were used to determined the expression of Hes1 at mRNA and protein levels in each generation of the cells.The proliferation and apoptosis of the BEP2D cells exposed to CSC were analyzed with the methods of MTT assay, flow cytometry and cell colony formation assay after treatment with Notch pathway bloker DAPT or liposome transfection with Hes1-siRNA.The expression of Hes1 in the peripheral small airway tissues of the smoking rats was evaluated by immunohistochemical staining.The expression of Hes1 in non-small-cell lung cancer and normal airway tissues was also detected by the methods of immunohistochemistry and RT-PCR.RESULTS: The BEP2D cells in the 70th generation had a malignant transformation phenotype.The expression of Hes1 in the BEP2D cells exposed to CSC for different time showed an increa-sing trend.DAPT and liposome transfection with Hes1-siRNA down-regulated the expression of Hes1, inhibited the cell proliferation and induced cell apoptosis.The expression of Hes1 in the airway mucosa of the rats exposed to cigarette smoke for 1 month and 6 months was significantly higher than that in control group.Cigarette smoking induced the expression of Hes1 in lung cancer and normal airway tissues.CONCLUSION: Hes1 may be involved in smoking-induced lung cancer by promoting the imbalance between apoptosis and proliferation.

Objective:To explore the application of failure mode and effects analysis (FMEA) in low-energy X-ray intraoperative radiotherapy (IORT), analyze its potential risks in IORT, and preliminarily explore the feasibility of FMEA in optimizing IORT management and reducing the occurrence of potential risks.Methods:An FMEA working group was established by the IORT team (1 radiologist, 1 radiology physicist, 2 surgeons, and 2 nurses) to apply the FMEA methodology to conduct a systematic risk assessment. The process modules were established, the potential failure modes and causes for each module were analyzed, the severity (SR), frequency of occurrence (OR) and likelihood of detection (DR) of failure modes were scored and the risk priority number (RPN) was calculated: RPN= SR × OR × DR. The possible errors and potential clinical impact of each part of the radiotherapy process were prospectively analyzed and understood, the causes and current measures were analyzed for each failure mode and preventive measures were proposed and risk management measures were taken accordingly.Results:The IORT process was divided into 8 modules with 14 failure modes. The highest OR value was unsatisfactory target area confirmation (7 points), the highest SR value was equipment failure to discharge the beam (10 points), the highest DR value was wrong key entry after dose calculation (7 points), the highest RPN values were unsatisfactory target area confirmation (210 points) and ineffective protection of endangered organs (180 points). Weaknesses were corrected according to priorities, workflows were optimized and more effective management methods were developed.Conclusion:FMEA is an effective method of IORT management and contributes to reducing the occurrence of potential risks.

Objective: To describe the prevalence of overweight and obesity among children and adolescents in Jinan City according the national and international criteria, as well as to compare the Chinese and international standards, so as to provide basic data for relevant intervention strategies. Methods: From September to December 2021, using the height and weight data obtained from the health checkup reports of 746 985 primary and secondary school students in Jinan City in 2021, body mass index(BMI) was calculated to evaluate the current status of the prevalence of overweight and obesity among this sample of children and adolescents aged 6- 18 years old. The consistency of the Chinese standard, U.S. Centers for Disease Control and Prevention (CDC) standard, International Obesity Task Force (IOTF) standard, and World Health Organization (WHO) standard was compared by calculating the weighted Kappa value. Results: Based on the Chinese, U.S. CDC, IOTF and WHO standards, the overweight rates of children and adolescents aged 6-18 years in Jinan were 17.3%, 16.5%, 21.1% and 13.6%, respectively, and the obesity rates were 23.1%, 19.5%, 13.5% and 6.6%, respectively. The overweight and obesity rates of boys were higher than those of girls under the four criteria (China: χ 2=1 418.48, 9 868.51, U.S. CDC: χ 2=145.78, 23 211.41, IOTF: χ 2=1 326.94, 13 615.62, WHO: χ 2=873.13, 46.41, P <0.01).The overweight rate of adolescents in the 13-18 year-old age group was higher than that of children aged 6-12 years (China: χ 2=29.13, U.S. CDC: χ 2=6.43, IOTF: χ 2=15.87, WHO: χ 2=19.48, P <0.01) and the obesity rate of the group aged 6-10 years was significantly higher than that of the group aged 11-18 years (China: χ 2=217.02, U.S. CDC: χ 2=227.54, IOTF: χ 2=171.91, WHO: χ 2=165.91, P <0.01). The rates of overweight and obesity in urban children and adolescents were slightly higher than those in rural children and adolescents (China: χ 2=25.06, 245.12, U.S. CDC: χ 2=56.13, 205.93, IOTF: χ 2= 182.34 , 22.04, WHO: χ 2=200.88, 21.61, P <0.01). The Chinese standard showed good agreement with the IOTF standard and U.S. CDC standard, with weighted Kappa values of 0.72 and 0.83, but poor agreement with the WHO standard, with a weighted Kappa value of 0.33. Conclusion The current prevalence of overweight and obesity among children and adolescents in Jinan is serious. The Chinese standard has good consistency with IOTF standard and American CDC standard, and poor consistency with WHO standard.

Objective To investigate the electrophysiological characteristics of hamster primary cortical neurons by using the whole-cell patch-clamp recordings in a Mg2+-free cell model of epilepsy.Methods The cerebral cortex were isolated for primary neuron culture from neonatal Syrian hamsters 1-2 days after birth.After 12 days,primary neurons were exposed to normal external(Mg2+group)or Mg2+-free external(Mg2+-free group)respectively for 3 hours,followed by a 24-hour incubation in normal culture medium.Excitatory postsynaptic currents(EPSC)and excitatory postsynaptic potentials(EPSP)were recorded in voltage clamp mode and current clamp mode using whole-cell patch-clamp recordings,respectively.Results Compared to Mg2+group,the frequency of EPSC[(124.38±75.15)Hz vs.(33.93±22.32)Hz,P<0.001](P<0.001)and EPSP[(37.05±38.37)Hz vs.(5.63±9.52)Hz,P<0.01]in primary cortical neurons from the Mg2+-free group increased significantly.Meanwhile,there were no statistically significant differences in the amplitude,area under the curve and half-width of EPSC and EPSP between the two groups(P>0.05).Conclusion The present study demonstrates an increase in excitability of primary hamsters cortical neurons after Mg2+depletion,suggesting that these neurons can be utilized to use to create cell models of epilepsy.