Objective:To investigate the role of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) in enamel development of mice.Methods:Single-cell RNA sequencing data of incisor tissues of postnatal day 7 (P7) mice and mandibular first molar tooth germs of P3.5 mice were used to analyze the expression of Wwp1 in dental epithelial cells. Immunohistochemistry was performed to observe the distribution and expression levels of WWP1 in the epithelium of mouse incisors and mandibular first molar tooth germs. Wwp1 knockout (Wwp1 KO) mice were generated and collected with their control littermates at P1, P7, three mice per group, as well as at P14, P28, 2 months (2M), and 3M, six mice per group. The enamel volumes of molars and incisors were analyzed using micro-CT. Scanning electron microscopy was employed to examine the enamel cross-sections of Wwp1 KO and control mice. Energy dispersive spectroscopy (EDS) was used to analyze the calcium and phosphorus content of the enamel rod of incisors. Immunofluorescence was performed to detect the expression of amelogenin (AMELX) in the ameloblasts of Wwp1 KO and control mice. Additionally, LS-8 ameloblast-like epithelial cells were cultured, and Wwp1 siRNA or overexpression plasmids were transfected to knock down or overexpress WWP1. The protein levels of AMELX were then assessed by Western blotting.Results:Single-cell sequencing result showed a high Wwp1 mRNA expression level in the epithelial cells of mouse incisors and mandibular molar tooth germs. Immunohistochemistry revealed the expression of WWP1 in presecretory, secretory, transitional, and mature ameloblasts. Wwp1 KO mice exhibited enamel developmental defects. The enamel volumes of molars and incisors in Wwp1 KO mice [(0.155±0.016), (0.300±0.017) μm 3] were reduced by 23.95% ( P<0.001) and 28.31% ( P<0.001) compared with the control group [(0.203±0.062), (0.418±0.023) μm 3] respectively. Scanning electron microscopy showed disorganized enamel structures in Wwp1 KO incisors and molars. EDS results showed the weight percent of calcium in the enamel rod of incisors decreased in Wwp1 KO mice [(20.74±0.91)%] compared with the control group [(30.30±3.83)%] ( P<0.001), and the calcium-to-phosphorus ratio decreased in Wwp1 KO mice (1.93±0.01) compared with the control group (2.02±0.01) ( P<0.001). Immunofluorescence showed weaker AMELX expression in ameloblasts of mandibular first molar tooth germs from P1 and P7 Wwp1 KO mice compared with the control group ( P<0.001, P<0.001). In LS-8 cells, Wwp1 knocked-down led to a decrease of AMELX protein expression, while WWP1 overexpression resulted in an increased AMELX protein level. Conclusions:WWP1 promotes ameloblast differentiation and enamel matrix mineralization, playing a critical role in enamel formation.

Objective:To investigate the role of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) in enamel development of mice.Methods:Single-cell RNA sequencing data of incisor tissues of postnatal day 7 (P7) mice and mandibular first molar tooth germs of P3.5 mice were used to analyze the expression of Wwp1 in dental epithelial cells. Immunohistochemistry was performed to observe the distribution and expression levels of WWP1 in the epithelium of mouse incisors and mandibular first molar tooth germs. Wwp1 knockout (Wwp1 KO) mice were generated and collected with their control littermates at P1, P7, three mice per group, as well as at P14, P28, 2 months (2M), and 3M, six mice per group. The enamel volumes of molars and incisors were analyzed using micro-CT. Scanning electron microscopy was employed to examine the enamel cross-sections of Wwp1 KO and control mice. Energy dispersive spectroscopy (EDS) was used to analyze the calcium and phosphorus content of the enamel rod of incisors. Immunofluorescence was performed to detect the expression of amelogenin (AMELX) in the ameloblasts of Wwp1 KO and control mice. Additionally, LS-8 ameloblast-like epithelial cells were cultured, and Wwp1 siRNA or overexpression plasmids were transfected to knock down or overexpress WWP1. The protein levels of AMELX were then assessed by Western blotting.Results:Single-cell sequencing result showed a high Wwp1 mRNA expression level in the epithelial cells of mouse incisors and mandibular molar tooth germs. Immunohistochemistry revealed the expression of WWP1 in presecretory, secretory, transitional, and mature ameloblasts. Wwp1 KO mice exhibited enamel developmental defects. The enamel volumes of molars and incisors in Wwp1 KO mice [(0.155±0.016), (0.300±0.017) μm 3] were reduced by 23.95% ( P<0.001) and 28.31% ( P<0.001) compared with the control group [(0.203±0.062), (0.418±0.023) μm 3] respectively. Scanning electron microscopy showed disorganized enamel structures in Wwp1 KO incisors and molars. EDS results showed the weight percent of calcium in the enamel rod of incisors decreased in Wwp1 KO mice [(20.74±0.91)%] compared with the control group [(30.30±3.83)%] ( P<0.001), and the calcium-to-phosphorus ratio decreased in Wwp1 KO mice (1.93±0.01) compared with the control group (2.02±0.01) ( P<0.001). Immunofluorescence showed weaker AMELX expression in ameloblasts of mandibular first molar tooth germs from P1 and P7 Wwp1 KO mice compared with the control group ( P<0.001, P<0.001). In LS-8 cells, Wwp1 knocked-down led to a decrease of AMELX protein expression, while WWP1 overexpression resulted in an increased AMELX protein level. Conclusions:WWP1 promotes ameloblast differentiation and enamel matrix mineralization, playing a critical role in enamel formation.

Objective:To evaluate the potential effects of serum lipid levels, appendicular skeletal muscle mass index (ASMI) and body mass index (BMI), together with its dynamic changes, on tumor progression in renal clear cell carcinoma patients, so as to inform body weight management.Methods:This prospective cohort study included a total of 100 patients with high-risk clear cell renal cell carcinoma. Serum lipid levels were detected, ASMI and BMI were measured using bioelectrical impedance analysis and the dynamic changes of BMI were tracked. The effects of BMI, ASMI and serum lipid levels on tumor progression within 2 years were explored.Results:Patients with normal BMI and low ASMI had 5.248 (95% CI: 1.946 to 14.153, P = 0.001) times higher risk of tumor progression than those who were overweight or obese. For every 0.1-unit increase in pre-operative HDL-C, the risk of tumor progression decreased by 0.771 (95% CI: 0.631 to 0.942, P = 0.011) times. Patients who experienced more than 5% decrease in BMI compared with baseline had 5.165 (95% CI: 1.735 to 15.370, P = 0.003) times the progression risk of patients whose BMI changed within ±5% from baseline. Conclusions:The advantage of obese clear cell carcinoma patients over normal-weight patients in tumor progression-free survival may be influenced by ASMI, pre-onset involuntary weight loss and lipid levels. Therefore, patient weight management should not merely focus on absolute BMI but tailor to individual characteristics, including cancer stage, body composition and metabolic status.

Objective:To explore the current status of clinical nurses' job well-being in Class Ⅲ Grade A general hospitals in Beijing, and analyze its influencing factors.Methods:From November 2019 to January 2020, convenience sampling method was used to select 528 clinical nurses from 4 ClassⅢ Grade A general hospitals in Beijing as the research object. The General Information Questionnaire, Perceived Organizational Support Scale, Psychological Capital Questionnaire-Revision (PCQ-R) and Employee Occupational Well-being Scale in Chinese Enterprises were used to conduct a cross-sectional survey to analyze the relationships among clinical nurses' perceived organizational support, psychological capital and job well-being and the influencing factors of well-being at work. A total of 528 questionnaires were collected in this survey, of which 518 were valid questionnaires, with an effective response rate of 98.1%.Results:Among 518 clinical nurses, the average score of job well-being was (5.13±0.96) . Pearson correlation analysis showed that the total scores of perceived organizational support and psychological capital of clinical nurses were positively correlated with the total score of job well-being with statistical differences ( r=0.802, 0.668; P<0.01) . Multiple linear regression analysis showed that monthly income, perceived organizational support and psychological capital were the influencing factors of clinical nurses' job well-being also with a statistical difference ( P<0.05) . Conclusions:Clinical nurses in four ClassⅢ Grade A general hospitals in Beijing have a good sense of job well-being. The more monthly income, the stronger the perceived organizational support, and the higher the psychological capital, the higher the job well-being of nurses is.

Objective To investigate the predictive value of serum albumin level in patients with severe sepsis .Methods One hundred and twenty cases of patients with severe sepsis admitted to Qilu Hospital ,Shandong University from April 2014 to October 2014 were prospectively enrolled .The serum albumin levels were measured and the laboratory and clinical data were collected at the onset of severe sepsis .Acute Physiology and Chronic Health Evaluation (APACHE ) Ⅱ score and Sequential Organ Failure Assessment (SOFA) score were calculated .Patients were grouped according to the prognosis by day 28 or stratified by albumin level . Prognostic factors were analyzed by multivariable Logistic regression .Results A total of 120 patients were enrolled with mean age of (57 .6 ± 18 .3) years ,among which 75 were male .The mean duration of hospitalization was (20 .1 ± 17 .8) days .The 28‐day mortality was 25 .8% (31/120) .The most common infection sources were respiratory tract (56 .7% ) ,abdominal/pelvis (19 .2% ) and bloodstream (9 .2% ) .Serum albumin level in survival group was significantly higher than that in death group ([32 .1 ± 6 .4] g/L vs [27 .5 ± 5 .5] g/L ,t=3 .562 ,P=0 .001) .Compared with survival group ,the patients in death group had higher APACHE Ⅱ and SOFA scores (22 .0 ± 9 .1 vs 13 .4 ± 7 .2;7 .1 ± 3 .7 vs 4 .3 ± 3 .5 ;t= —5 .372 and —3 .690 ,both P<0 .05) .Along with the decrease of serum albumin level ,the incidence of bloodstream infection ,solid tumor ,septic shock ,acute kidney injury and liver injury significantly increased .Patients with lower albumin level had significantly higher SOFA scores and 28‐day mortality (all P<0 .05) .Multivariable regression analysis showed that albumin level lower than 28 g/L and higher APACHE Ⅱ score were independent risk factors for mortality (OR=4 .156 ,95% CI:1 .198—14 .415 ;OR=1 .121 ,95% CI:1 .039—1 .210;both P<0 .05) .Conclusions A significantly lowered serum albumin level would increase the risk of mortality in patients with severe sepsis .The combination of albumin level and APAHCE Ⅱ score might be beneficial to evaluate the prognosis .