Objective:To observe the effects of lactoferrin(LF)on the proliferation,migration and osteogenic differentiation of human periodontal ligament cells(HPDLCs)in vitro.Methods:HPDLCs were cultured and identified.The proliferation and migration of HP-DLCs cultured with 0,10 and 20 μg/ml lactoferrin respectively were tested by MTT assay,Transwell assay and scratch test.The os-teogenic differentiation of the cells was evaluated using alizarin red staining and real-time PCR.Results:Lactoferrin at 10 and 20μg/ml increased the proliferation(P <0.05),increased the quantities of mineralized nodules and the expression of alkaline phospha-tase(ALP),osteocalcin(OCN)and osteopontin(OPN)(P <0.05).Conclusion:Lactoferrin promotes the proliferation,migration and osteogenic differentiation of HPDLCs.

Objective To investigate the influence of“double low”technology(low concentration iodinated contrast agent and low-dose scan) combined with body mass index(BMI) on radiation dose and image quality of contrast-enhanced upper abdominal CT examination. Methods One hundred and twenty patients who received upper abdominal enhanced CT examination were randomly divided into 4 groups:group A1, the iodinated contrast agent iodixanol(270 mg/ml), BMI<18.5 kg/m2 and 80 kVp;group A2, the iodinated contrast agent iodixanol(270 mg/ml), 18.5 kg/m2≤BMI≤24.9 kg/m2 and 100 kVp; group B1, the iodinated contrast agent ioversol(320 mg/ml),<18.5 kg/m2 and 120 kVp; group B2, the iodinated contrast agent ioversol(320 mg/ml), 18.5 kg/m2≤BMI≤24.9 kg/m2 and 120 kVp. Image quality was subjectively scored, the objective parameters(noise, CT values of abdominal aorta and liver parenchyma, contrast noise ratio of abdominal aorta and liver parenchyma) were evaluated and radiation dose was recorded. The differences of the indexes between A1 and B1 groups, A2 and B2 groups were compared with Mann-Whitney U test and pared-samples t test. Results All CT images were good. No images with 4 scores were obtained. No significant difference was found between group A1 and B1, between group A2 and B2(P>0.05). There was no significant difference in contrast noise ratio of liver parenchyma(P>0.05), while significant differences existed in CT values of abdominal aorta and liver parenchyma, contrast noise ratio of abdominal aorta between group A1 and B1(P<0.05). Significant differences existed in the parameters above mentioned between group A2 and B2, respectively(P<0.05). Radiation dose was lower in group A1 than in group B1 and in group A2 than in group B2(P<0.05), respectively. Radiation dose was decreased by 40.1%(0.89/2.22) in group A1 than group B1 while radiation dose decreased by 56.9%(3.02/5.31) in group A2 than group B2. Conclusion According to BMI, the low concentration iodinated contrast agent and low-dose scan CT scanning technology could effectively reduce radiation dose and generate ideal images during the contrast-enhanced upper abdominal CT examination.

ObjectiveTo study the effect of anticoagulant therapy with low molecular weight heparin (LMWH) on coagulation and inflammation markers in sepsis patients.Methods A prospective randomized controlled trial was conducted. Sixty sepsis patients admitted to intensive care unit (ICU) of Zhengzhou University People's Hospital from March 2012 to May 2014 were divided into control group and observation group, with 30 cases in each group. The observations were begun as soon as the diagnosis of sepsis was established, and the observation time was 7 days. All sepsis patients were treated according to the 2008 international sepsis treatment guidelines. Every patient in the observation group was subcutaneously injected with LMWH 0.6 mL on the first day of ICU admission, twice a day for 7 days. The blood from peripheral vein was collected at 1, 3, 5, 7 days of treatment, and CD62p, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), D-dimmer was determined by immunoturbidimetry, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score was recorded, and incidence of multiple organ dysfunction syndrome (MODS) was also evaluated.Results There were no significant differences in values of all parameters, including CD62p, D-dimmer, IL-6, TNF-α, and APACHEⅡ score at 1 day of treatment. The values of all parameters in observation group were gradually decreased. CD62p at 3 days of treatment and D-dimmer, IL-6, TNF-α, and APACHEⅡ score at 5 days of treatment were significantly lower than those at 1 day of treatment. The values in the control group were decreased at first and then increased, as D-dimmer, IL-6 and TNF-α were significantly higher on the 5th day than those at 1 day of treatment. Compared with control group, CD62p, D-dimmer, IL-6, TNF-α and APACHEⅡ score on the 7th day of treatment were significantly lowered in observation group [CD62 (μg/L): 22.64±2.88 vs. 31.52±2.81, D-dimmer (g/L): 1.32±0.46 vs. 4.79±0.82, IL-6 (ng/L): 5.84±1.87 vs. 49.64±3.12, TNF-α (ng/L): 21.04±3.15 vs. 130.58±6.26, APACHEⅡ score: 9.71±2.02 vs. 14.17±2.38, allP< 0.05]. Correlation analysis showed that in observation group, CD62p, D-dimmer, IL-6, and TNF-α were positively correlated with APACHEⅡ score (r value was 0.907, 0.868, 0.880, 0.693, respectively, all P=0.000). The incidence of MODS in observation group was significantly lower than that in the control group [26.7% (8/30) vs. 46.7% (14/30),χ2=3.943,P= 0.028].Conclusions LMWH, which was given early in sepsis, can significantly down-regulate the expression of CD62p, D-dimmer, IL-6 and TNF-α, and reduce the incidence of MODS. Some indicators regarding coagulation and inflammation can be used as supplementary indicators to severity scores, and it may be able to improve the accuracy of scoring systems for sepsis.

Objective:To evaluate the role of B/glycogen synthase kinase-3β (Akt/GSK-3β) signaling pathway in interleukin-4 (IL-4)-induced reduction of cerebral ischemia-reperfusion (I/R) injury in mice and the relationship with autophagy.Methods:Forty clean-grade healthy male Balb/c mice, aged 10-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), cerebral I/R group (group IR), IR plus IL-4 group, and IR plus IL-4 plus Akt inhibitor LY294002 group (IR+ IL-4+ LY group). Cerebral I/R was induced by 60 min middle cerebral artery occlusion followed by 24 reperfusion in anesthetized mice.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model in group IL-4.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model, and LY294002 15 nmol/kg was simultaneously injected via the tail vein in group IR+ IL-4+ LY.Neurological function was assessed and scored at 24 h of reperfusion, and then animals were sacrificed and brains removed for determination of cerebral infarct size (by TTC assay), cell apoptosis, autophagosome count (with transmission electron microscope), levels of superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) (using colorimetric assay), phosphorylation of Akt and GSK-3β, expression of LC3 and Beclin-1 (by Western blot). The apoptosis index and LC3Ⅱ/LC3Ⅰ ratio were calculated. Results:Compared with Sham group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in the other three groups ( P<0.05). Compared with IR group, the neurological scores, cerebral infarct size and apoptosis index were significantly decreased, the SOD activity in brain tissues was increased, levels of MDA and ROS were decreased, phosphorylation of Akt and GSK-3β was increased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were decreased in IR+ IL-4 group ( P<0.05), and no significant change was found in the parameters mentioned above in IR+ IL-4+ LY group ( P>0.05). Compared with IR+ IL-4 group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in IR+ IL-4+ LY group ( P<0.05). Conclusion:IL-4 can inhibit cell autophagy through activating Akt/GSK-3β signaling pathway and thus reduces cerebral I/R injury in mice.

Objective:To evaluate the relationship between the second messenger cyclic GMP-AMP (cGAS)-cyclic GMP-AMP receptor stimulator of interferon genes (STING) signaling pathway and ferritinophagy in the early stage of cerebral ischemia-reperfusion (I/R) in mice.Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 21-25 g, were divided into 4 groups ( n=6 each) using a random number table method: sham group, cerebral I/R injury group (CIRI group), cerebral I/R injury + cGAS inhibitor group (CIRI + RU group), and cerebral I/R injury + cGAS inhibitor + overexpressed nuclear receptor coactivator 4 (NCOA4) group (MCAO + RU + LV-NCOA4 group). The model of cerebral I/R injury was developed using the middle cerebral artery occlusion (MCAO) in anesthetized animals.In CIRI+ RU group, cGAS inhibitor 5 mg/kg was intraperitoneally injected at 10 min before reperfusion.In CIRI+ RU+ LV-NCOA4 group, NCOA4-overexpressing lentivirus (1×10 9 TU/ml) 2 μl was injected into the ventricle at 7 days before MCAO, and the other operations were the same as those previously described in CIRI+ RU group.After 6 h of reperfusion, the neurological function deficits were assessed and scored, then the mice were sacrificed, and brains were removed for determination of the cerebral infarct size (by TTC method), MDA content (by TBA method), activity of SOD (by WST-1 method), and expression of cGAS, STING, NCOA4, ferritin, and microtubule-associated protein 1 light chain 3B (LC3B) (by Western blot). Results:Compared with Sham group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05). Compared with CIRI group, the neurological function deficit score and cerebral infarct size were significantly decreased, SOD activity was increased, MDA content was decreased, the expression of cGAS, STING, NCOA4 and LC3B was down-regulated, and the expression of ferritin was up-regulated in CIRI+ RU group ( P<0.05). Compared with CIRI+ RU group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05), and no significant change was found in the expression of cGAS and STING in CIRI+ RU+ LV-NCOA4 group ( P>0.05). Conclusions:The cGAS-STING signaling pathway can promote the over-activation of ferritinophagy, enhance oxidative stress, and thus induce early CIRI in mice.

Objective The SEER database was utilized to analyze the prognosis and related factors for patients with locally resectable esophageal cancer undergoing preoperative chemoradiotherapy.A nomogram for predicting survival was established to provide reference for screening patients receiving preoperative chemoradiotherapy.Methods Patients diagnosed with stage T1b-4aN0-3M0(7th version AJCC in 2010) resectable esophageal cancer receiving preoperative chemoradiotherapy between 2010 and 2015 were selected from the SEER database.The survival rate was determined by Kaplan-Meier method.The univariate analysis was performed by log-rank test.The multivariate analysis was conducted by Cox test.The nomogram for survival prediction was established by using R software.The predicting accuracy of the nomogram was evaluated by C-index and calibration curve.Results A total of 1 697 eligible patients were included.Univariate analysis showed that sex,T stage,N stage and tumor differentiation were significantly associated with overall survival (OS) and cancer-specific survival rate (CSS)(all P＜0.001),and age (P=0.027) was significantly correlated with OS.Multivariate analysis demonstrated that age,sex,tumor differentiation and N stage were significantly associated with OS.Sex,tumor differentiation,T stage and N stage were significantly correlated with CSS (all P＜0.05).After the prognostic factors were included into the nomogram,the C-index for 5-year OS and CSS was 0.60 and 0.61.The model for predicting survival of patients with esophageal squamous cell cancer was established by using the same method.The C-index for the OS and CSS was 0.62 and 0.64.Conclusions Sex,clinical stage and tumor differentiation are prognostic factors of CSS in patients with locally resectable esophageal cancer undergoing preoperative chemoradiotherapy followed by surgery.The nomogram established according to the data above can provide certain reference for the selection of preoperative chemoradiotherapy combined with surgery.

Objective:To investigate the effect of homogeneous management combined with staged teaching on physicians receiving standardized training of hepatobiliary surgery.Methods:A total of 46 physicians who received standardized training in Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, from January to March 2020 were selected as control group and were given conventional teaching, and 50 physicians who received standardized training from April to June 2020 were selected as observation group and were given homogeneous management combined with staged teaching. The two groups were compared in terms of professional level, clinical ability, and the degree of satisfaction with teaching before and after teaching. SPSS 24.0 was used to perform the independent samples t-test, the paired t-test, the chi-square test, and the rank sum test. Results:After teaching, both groups had significant increases in the scores of theoretical examination and operation skill examination, and compared with the control group, the observation group had significantly higher scores of theoretical examination (94.57±3.28 vs. 90.32±2.12) and operation skill examination (94.37±4.18 vs. 91.25±3.46). After teaching, both groups had significant increases in the scores of clinical consultation, physical examination, humanistic concern, clinical diagnosis, communication ability, organizational ability, and overall evaluation, and the observation group had significantly higher scores of the above seven aspects than the control group (6.98±0.94/6.45±0.14/6.95±0.88/6.65±0.93/6.53±0.26/6.84±0.92/6.58±0.35 vs. 6.13±0.31/6.21±0.76/6.21±0.42/6.18±0.35/6.32±0.61/6.33±0.24/6.25±0.71). The observation group had a significantly higher overall satisfaction rate than the control group [94.00% (47/50) vs. 78.26% (36/46)].Conclusion:In the standardized training and teaching of hepatobiliary surgery, homogeneous management combined with staged teaching can improve the professional level and clinical ability of physicians and enhance the degree of satisfaction with teaching.

Objective:To explore the differences of risk stratification of very high-risk or extreme high-risk atherosclerotic cardiovascular diseases (ASCVD) and the attainment rates of low-density lipoprotein cholesterol (LDL-C) management targets evaluated by three different criteria, and the causal attributions of these differences.Methods:Patients with ASCVD were consecutively enrolled from January 1 to December 31 in 2019, and were evaluated for very high-risk or extreme high-risk and LDL-C goal attainment rates with 2018 American guideline on the management of blood cholesterol (2018AG), 2019 China Cholesterol Education Program (CCEP) Expert Advice for the management of dyslipidemias (2019EA) and 2020 Chinese expert consensus on lipid management of very high-risk ASCVD patients(2020EC), respectively. The causal attributions of the differences in attainment rates were analyzed as well.Results:A total of 1 864 ASCVD patients were included in this study. According to 2018AG, 2019EA and 2020EC, the proportions of the patients with very high-risk or extreme high-risk were 59.4%, 90.7%, and 65.6%, respectively. The absolute LDL-C target attainment rates were 37.2%, 15.7%, and 13.7%, respectively, the differences between each two rates were statistically significant (all P<0.001). As to the differences in attainment rates between 2020EC and 2018AG, 61.5% were due to the different LDL-C goal attainment values and 38.5% were caused by the different risk stratifications, while for the differences between 2020EC and 2019EA attainment rates, different LDL-C goal attainment values were responsible for 13.2%, and different risk stratifications were responsible for 86.8% of the differences. Conclusions:There are significant differences in the proportions and LDL-C attainment rates among the three different criteria for very high-risk or extreme high-risk ASCVD. 2020EC showed a moderate proportion of patients with extreme high-risk, and had the lowest LDL-C attainment rate. The differences between 2020EC and 2018AG are mainly due to the LDL-C target values, and the differences between 2020EC and 2019EA are mainly caused by the risk stratifications.

Development of thoracolumbar vertebra (TLV) and rib primordium (RP) is a common evolutionary feature across vertebrates, although whole-organism analysis of the expression dynamics of TLV- and RP-related genes has been lacking. Here, we investigated the single-cell transcriptome landscape of thoracic vertebra (TV), lumbar vertebra (LV), and RP cells from a pig embryo at 27 days post-fertilization (dpf) and identified six cell types with distinct gene expression signatures. In-depth dissection of the gene expression dynamics and RNA velocity revealed a coupled process of osteogenesis and angiogenesis during TLV and RP development. Further analysis of cell type-specific and strand-specific expression uncovered the extremely high level of HOXA10 3'-UTR sequence specific to osteoblasts of LV cells, which may function as anti-HOXA10-antisense by counteracting the HOXA10-antisense effect to determine TLV transition. Thus, this work provides a valuable resource for understanding embryonic osteogenesis and angiogenesis underlying vertebrate TLV and RP development at the cell type-specific resolution, which serves as a comprehensive view on the transcriptional profile of animal embryo development.