ObjectiveTo study the effect of Qizhu Kang'ai prescription （QZAP） on the gluconeogenesis enzyme phosphoenolpyruvate carboxykinase 1 （PCK1） in the liver of mouse model of liver cancer induced by diethylnitrosamine （DEN） combined with carbon tetrachloride （CCl₄） and Huh7 cells of human liver cancer， so as to explore the mechanism on regulating metabolic reprogramming and inhibiting cell proliferation of liver cancer cells. MethodDEN combined with CCl₄ was used to construct a mouse model of liver cancer via intraperitoneal injection. A normal group， a model group， and a QZAP group were set up， in which QZAP （3.51 g·kg^-1） or an equal volume of normal saline was administered daily by gavage， respectively. Serum and liver samples were collected after eight weeks of intervention. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， γ-glutamyltransferase （γ-GT）， and alpha-fetoprotein （AFP） in mice were detected to evaluate liver function changes of mice in each group. Hematoxylin-eosin （HE） staining and Sirius red staining were used to observe pathological changes in liver tissue. In the cell experiment， Huh7 cells were divided into blank group， QZAP low， medium， and high dose groups and/or PCK1 inhibitor （SKF-34288 hydrochloride） group， and Sorafenib group. The corresponding drug-containing serum and drug treatment were given， respectively. Cell counting kit-8 （CCK-8） method， colony formation experiment， Edu fluorescent labeling detection， intracellular adenosine triphosphate （ATP） content detection， and cell cycle flow cytometry detection were used to evaluate the proliferation ability， energy metabolism changes， and change in the cell cycle of Huh7 cells in each group. Western blot was used to detect the protein expression levels of PCK1， serine/threonine kinase （Akt）， phosphorylated Akt （p-Akt）， and cell cycle-dependent protein kinase inhibitor 1A （p21）. ResultCompared with the model group， the pathological changes such as cell atypia， necrosis， and collagen fiber deposition in liver cancer tissue of mice in the QZAP group were alleviated， and the number of liver tumors was reduced （P<0.01）. The serum ALT， AST， γ-GT， and AFP levels were reduced （P<0.01）. At the cell level， compared with the blank group， low， medium， and high-dose groups of QZAP-containing serum and the Sorafenib group could significantly reduce the survival rate of Huh7 cells （P<0.01） and the number of positive cells with Edu labeling （P<0.01） and inhibit clonal proliferation ability （P<0.01）. The QZAP groups could also reduce the intracellular ATP content （P<0.05） and increase the distribution ratio of the G₀/G₁ phase of the cell cycle （P<0.05） in a dose-dependent manner. Compared with the model group and blank group， PCK1 and p21 protein levels of mouse liver cancer tissue and Huh7 cells in the QZAP groups were significantly reduced （P<0.05，P<0.01）， and the p-Akt protein level was significantly increased （P<0.01）. Compared with the blank group， the ATP content and cell survival rate of Huh7 cells in the SKF-34288 hydrochloride group were significantly increased （P<0.05）， but there was no statistical difference in the ratio of Edu-positive cells and the proportion of G₀/G₁ phase distribution. Compared with the SKF-34288 hydrochloride group， the QZAP combined with the SKF-34288 hydrochloride group significantly reduced the ATP content， cell survival rate， and Edu-positive cell ratio of Huh7 cells （P<0.05） and significantly increased the G₀/G₁ phase distribution proportion （P<0.05）. ConclusionQZAP may induce the metabolic reprogramming of liver cancer cells by activating PCK1 to promote Akt/p21-mediated tumor suppression， thereby exerting an anti-hepatocellular carcinoma proliferation mechanism.

The exacerbation of progressive multiple sclerosis (MS) is closely associated with obstruction of the differentiation of oligodendrocyte progenitor cells (OPCs). To discover novel therapeutic compounds for enhancing remyelination by endogenous OPCs, we screened for myelin basic protein expression using cultured rat OPCs and a library of small-molecule compounds. One of the most effective drugs was pinocembrin, which remarkably promoted OPC differentiation and maturation without affecting cell proliferation and survival. Based on these in vitro effects, we further assessed the therapeutic effects of pinocembrin in animal models of demyelinating diseases. We demonstrated that pinocembrin significantly ameliorated the progression of experimental autoimmune encephalomyelitis (EAE) and enhanced the repair of demyelination in lysolectin-induced lesions. Further studies indicated that pinocembrin increased the phosphorylation level of mammalian target of rapamycin (mTOR). Taken together, our results demonstrated that pinocembrin promotes OPC differentiation and remyelination through the phosphorylated mTOR pathway, and suggest a novel therapeutic prospect for this natural flavonoid product in treating demyelinating diseases.
Animals ; Cell Differentiation ; Flavanones ; Mice ; Mice, Inbred C57BL ; Myelin Sheath/metabolism* ; Oligodendroglia/metabolism* ; Rats ; Remyelination ; Signal Transduction ; TOR Serine-Threonine Kinases/metabolism*

The exacerbation of progressive multiple sclerosis (MS) is closely associated with obstruction of the differentiation of oligodendrocyte progenitor cells (OPCs). To discover novel therapeutic compounds for enhancing remyelination by endogenous OPCs, we screened for myelin basic protein expression using cultured rat OPCs and a library of small-molecule compounds. One of the most effective drugs was pinocembrin, which remarkably promoted OPC differentiation and maturation without affecting cell proliferation and survival. Based on these in vitro effects, we further assessed the therapeutic effects of pinocembrin in animal models of demyelinating diseases. We demonstrated that pinocembrin significantly ameliorated the progression of experimental autoimmune encephalomyelitis (EAE) and enhanced the repair of demyelination in lysolectin-induced lesions. Further studies indicated that pinocembrin increased the phosphorylation level of mammalian target of rapamycin (mTOR). Taken together, our results demonstrated that pinocembrin promotes OPC differentiation and remyelination through the phosphorylated mTOR pathway, and suggest a novel therapeutic prospect for this natural flavonoid product in treating demyelinating diseases.

Objective:To construct a standardized training course system for new nurses in radiology department based on their post competency.Methods:The first draft of standardized training course for new nurses in radiology department was drawn up by referring to domestic and foreign literature, expert interview and group discussion. 20 experts in radiology department from 12 third grade hospitals were consulted for two rounds by Delphi method, and the index weight at all levels was determined by AHP.Results:The standardized training course system of new nurses in radiology department based on post competency includes four first level indicators (knowledge, skills, situational decision-making, humanistic quality), 10 second level indicators and 55 third level indicators. In the second round of expert consultation, the positive coefficient of experts was 85%, 100% and the authoritative coefficient was 0.75 and 0.80 respectively; In the second round expert consultation, the harmony coefficient of the first, second and third level indexes was 0.401 and 0.493 respectively, P < 0.01. Conclusion:the standardized training course system of new nurses in radiology department based on post competency is scientific and reasonable, which can provide reference for standardized training of new nurses in radiology department.

Objective:To investigate clinical significance and related factors of magnetic resonance hyperintense vessel sign (HVS).Methods:The clinical data and related imaging parameters of 109 patients with acute anterior circulation occlusion cerebral infarction, who admitted to Northern Theater Command General Hospital of People′s Liberation Army from April 2017 to August 2019, were analyzed retrospectively. Brain magnetic resonance imaging (MRI) examinations including fluid attenuated inversion recovery (FLAIR), diffusion weighted imaging (DWI) and three dimensional time of flight magnetic resonance angiography (3D TOF MRA) sequences within 24 hours of onset were performed. According to the distribution range of HVS in FLAIR sequence, the patients were divided into four grades (0, 1, 2 and 3), grades 0 and 1 belonging to HVS low grade group, and grades 2 and 3 HVS high grade group. Univariate and multivariate analyses were made to explore related factors of HVS. Fifty-two patients who completed baseline CT within six hours of onset before MRI examination were performed CT-Alberta Stroke Program Early CT Score (CT-ASPECTS) and DWI-Alberta Stroke Program Early CT Score (DWI-ASPECTS).The difference between CT-ASPECTS and DWI-ASPECTS was calculated. When the difference of ASPECTS ≤1, they were categorized as ASPECTS unchanged group (AN group); when the difference of ASPECTS>1, they were categorized as ASPECTS changed group (AY group). These two groups were compared to explore whether there was any difference in HVS grade, and Spearman correlation analysis was performed to investigate the relationship between HVS grade and the difference of ASPECTS.Results:The difference of hyperlipidemia, TOAST classification (large artery atherosclerosis (LAA), other etiology (SOE) or undetermined etiology (SUE)) and Willis circle classification (types Ⅰ, Ⅱ, Ⅲ and Ⅳ) between HVS groups were remarkable (58.6% (34/58) vs 37.3% (19/51), χ2=4.959, P=0.026; 23/5/23 vs 43/1/14, P=0.004; 3/14/12/22 vs 7/29/14/8, χ2=13.124, P=0.004). Other clinical factors and the locations of vessel occlusion did not show significant difference ( P>0.05). Multivariate Logistic regression analysis indicated that LAA in TOAST classification (LAA vs SOE or SUE, OR=3.054, 95% CI1.257-7.422, P=0.014), Willis circle type Ⅰ (type Ⅰ vs type Ⅳ, OR=5.494, 95% CI1.074-28.091, P=0.041), and type Ⅱ (type Ⅱ vs type Ⅳ, OR=5.571, 95% CI1.895-16.372, P=0.002) were independent related factors to stimulate wide distribution of HVS. The grades of HVS were significantly different between the AN group and the AY group (1/15 vs18/18, χ2=9.114, P=0.002). Spearman correlation analysis showed that HVS grade was negatively correlated with the difference of ASPECTS ( r=-0.573, P<0.001). Conclusions:Both TOAST and Willis circle classifications are crucial factors affecting HVS distribution. HVS distribution range reflects the status of collateral compensatory. Recognizing HVS may help to evaluate the progress of early cerebral infarction volume.

Objective: To explore the diagnosis value of late gadolinium enhancement(LGE) detected by magnetic resonance imaging(MRI) in acute myocardial infarction(AMI) patients. Methods: The clinical and MRI data of 52 AMI patients hospitalized from January 2016 to July 2017 in our hospital were retrospectively analyzed. All patients received medication and revascularization therapies after admission and cardiac magnetic resonance examination was performed within 1 week after admission. According to whether there was LGE, AMI patients were divided into LGE(+) group(33 cases) and LGE(-) group(19 cases). According to the existence of microvascular obstruction(MVO) and/or intramyocardial hemorrhage(IMH),LGE(+) patients were divided into MVO/IMH(+) group(18 cases) and MVO/IMH(-) group(15 cases). Results: (1)There were no statistical significance between the LGE(+)group and LGE(-)group in the age, gender,smoking history, hypertension, diabetes mellitus, dyslipidemia, ventricular arrhythmia, culprit vessel, left ventricular end-diastolic volume(LVEDV), and left ventricular end-systolic volume(LVESV) (all P>0.05). The left ventricular ejection fraction was significantly lower in LGE(+) group than in LGE(-) group( (38.7±17.6) % vs. (51.2±7.9)%, P=0.001). (2)The infarct size was positively correlated with LVEDV and LVESV(r=0.436,P=0.011;r=0.479,P=0.005,respectively), and negatively correlated with left ventricular ejection fraction (r=-0.641, P<0.001) in LGE(+) group. (3) The infarct size, LVEDV, and LVESV were significantly higher in MVO/IMH(+) group thanin MVO/IMH(-) group ((26.5±7.3)%vs. (16.2±8.3)%, P=0.001; (145.7±40.9)ml vs. (112.2±23.8)ml,P=0.009; (90.0±30.8)ml vs. (61.4±19.0)ml,P=0.004, respectively), and the left ventricular ejection fraction was significantly lower in MVO/IMH(+) group than in MVO/IMH(-) group ((29.8±15.0)% vs. (49.3±14.5)%, P=0.001). Conclusions LGE detected bycardiac magnetic resonance can provide useful information on the myocardial necrosis extent of AMI patients. Presence of MVO/IMH in LGE(+) patients is linked with larger infarct size and worse cardiac dysfunction in AMI patients.

Objective To evaluate the opening level and optimal time window of the blood-brain barrier induced by adenosine A2 receptor agonist ( Lexiscan) via dynamic enhanced MRI. Methods Twen-ty New Zealand white rabbits were divided into experiment group ( group A, n=10) and control group ( group B, n=10) . Rabbits in group A were injected with Lexiscan and rabbits in group B were injected with physiological salt via ear vein, then the coronary scanning was performed. Contrast enhanced MRI was performed at different time points ( 5, 10, 15, 20 min, and then every 10 min, until 2 h) following the in-fusion of Gd-diethylene triamine pentaacetic acid (DTPA). The signal intensity (SI) of region of interest ( ROI) was measured and the percent enhancement of SI was calculated. Evens blue staining results in brain tissues were observed. Pair t test was used to analyze the data. Results The percent enhancement of SI in group A significantly increased to (40. 93±3.70)％ at 5 min, reached the maximum of (43.03±3.62)％ at 30 min, slowly decreased until 50 min, and got to a stable level at almost 80 min. At each time point, the per-cent enhancement of SI in group A was significantly higher than that in group B ( t values:6.88-20.28, all P<0. 05) . The staining was evident in group A. Conclusions Lexiscan can open blood-brain barrier tem-porarily and reversibly, and the optimal opening time window is 10-50 min post-injection.

In this study, a multiplex RT-PCR method was developed for detection of seven diarrhea-associated porcine viruses, including porcine teschovirus (PTV), porcine sapovirus (PSV), porcine deltacornavirus (PDCoV), porcine kobuvirus (PKV), porcine sapovirus (PSaV), porcine astrovirus (PAstV) and porcine torovirus (PToV). A total of 419 samples were screened by this method and results showed that PKV had the highest positive rate of 26.98%?45.79% and its mixed infection rate reached 9.52%-18.54%. On account of high positive rate of PKV and its important role in diarrhea disease, complete genomic sequences of three PKV positive samples were further sequenced. Three PKV labeled as PD-PKV, JS-PKV and CM-PKV were classified into porcine kobuvirus genus and had far genetic distance with other kobuviruses. The complete genome homologies among them were 88.1%-89.1%. CM-PKV had the highest identity with the Chinese strain JS-02a-CHN/2013 reported in 2013 while JS-PKV and PD-PKV were most closed to the K-30-HUN/2008/HUN strain reported in Hungary in 2008. This illustrates the significant genetic differences of the different PKV isolates in Shanghai while its relationship with the viral pathogenicity still needs to be explored. This research provides references for further understanding the prevalence of PKV and its role in swine diarrhea.

Background Light-induced retinal damage results in the damage of retinl pigment epithelial (RPE) cells and therefore affects the pathogenesis and development of age-related macular degeneration (AMD).Studies showed that tissue factor (TF) is overexpressed in oxidative damaged RPE cells and the choroidal neovascularization (CNV) of AMD,speculating that the suppression of TF can prevent the damage of RPE cells and inhibit CNV.Objective This study was conducted to observe the protective effects of TF targeting peptide (TFTP),a new drug of autologous synthesis,on human RPE-cells induced by blue light.Methods Human RPE cells were isolated from donor eye and cultured.Cultured cells were divided into blank control group,model group and TFTP treated group.Light-induced RPE cell damage model was established by exposuring the cells in the blue light of (4.0±-0.5) mW/cm2 for 12 hours in the model group,and different concentrations (10,100,150,200,300 μmol/L) of TF-TP were added into the medium to pretreat the cells for 24 hours and then exposed the cells to the blue light for 12 hours in the TF-TP groups.The cell viability was determined by CCK-8 assay.The morphology and ultrastructure in the cells were observed under the inverted microscope and transmission electron microscope.The apoptosis of the cells was assayed by Hoechst staining.The expressions of TF and apoptosis-related protein bax,bcl-2 in the cells were determined by Western blot.Results CCK-8 assay showed that there was no significant difference in the cell viability among blank control group and different concentrations TF-TP groups (F=2.15,P =0.11).The cell survival rate of blank control group,model group and 150 μmol/L TF-TP group was (100.0±0.00) ％,(43.79±6.55) ％ and (63.45±3.57) ％,and the survial rate was increased in the 150 μmol/L TF-TP group compared with the model group (P =0.00),and 150 μmol/L was detemined as a optimal concentration of TF-TP.A lot of shrinkage,deformation,suspension cells were exhibited under the optical microscope,and decrease of microvilli structure,rupture of mitochondrial cristae and vacuolar degeneration of the cells were found in the model group,and the damage of the cells were evidently lightened in the 150 μ mol/L TF-TP group.The apoptosis rate of the cells were (0.98 ±0.19)％,(9.98 ±0.82) ％ and (5.73 ±0.88) ％ in the blank group,model group and 150 μmol/L TF-TP group,respectively,with a significant difference among the groups (F =206.18,P =0.00),and the apoptosis rate of the cells in the 150 μmol/L TF-TP group was significantly lower than that in the model group (P＜0.05).Compared with the blank control group,the relative expression of bax and TF was obviously increased and that of bcl-2 was decreased in the model group;while the expression of bax and TF was lower,and that of bcl-2 was higher in the 150 μmol/L TF-TP group compared with the model group (all at P ＜ 0.05).Conclusions Pretreation of TF-TP can lessen cell apoptosis and increase cell survival rate and therefore plays a protective role to blue light-induced human RPE cells possibly by inhibiting bax/bcl-2 apoptotic pathways mediated by TF.

Objective To analyze the effects of quality supervision and continuous improvement system on optimizing in-hospital diagnosis and treatment process in patients with acute ischemic stroke (AIS).Methods From September 2013 to May 2016,424 consecutive patients with AIS treated with intravenous thrombolysis and/or endovascular therapy in Changhai Hospital,the Second Military Medical University were enrolled retrospectively.They were analyzed according to the annual running process (the first year[from September 2013 to August 2014],the second year[from September 2014 to August 2015],and the third year[from September 2015 to May 2016]).The spend time and delay (DTN>60 min,DTP>90 min) rate of each treatment process in the first,second,and third year (time from door-to-imaging[DTI],door-to-needle[DTN],imaging-to-needle (ITN),door-to-groin puncture (DTP) and imaging-to-groin puncture (ITP) were compared.Taking the time periods (>median) of having significant differences of the spend time of the treatment processes as the dependent variables in the first,second,and third year,the influence of the years and treatment modalities on delay was observed.The difference of constituent ratio of the reasons for delay in intravenous thrombolysis and endovascular therapy (objective reasons/other reasons) in different years were analyzed.Results (1) DTIs were 23.0 (11.0,42.0) min,22.0 (10.1,39.0) min,and 13.0 (6.0,27.0) min,respectively,and DTNs were 50.0 (30.0,77.1) min,45.0 (30.0,70.2) min,and 36.0 (24.0,57.0) min,respectively in the first,second,and third year.The spending time was shortened year by year.There were significant differences among the different years (all P<0.01).The spending time of DTP had a tendency to be shortened,but there were significant differences among different years (P=0.06).There were no significant differences between the spending time of ITN and ITP (all P>0.05).(2) The DTN delay rates were 33.3% (40/120),20.7% (29/140),and 8.1% (9/111),respectively in the first,second,and third year.There were significant differences among the 3 years (x2=22.111,P<0.01).There were no significant differences among the DTP delay rates (P=0.08).(3) Multivariate Logistic regression analysis showed that taking the first years as a reference,the risk of DTI delay was reduced in the third year (OR,0.174,95%CI 0.101-0.298,P<0.01),the risks of DTN delay were reduced in the second and third year (OR,0.564,95%CI 0.338-0.941;OR,0.180,95%CI 0.101-0.320,all P<0.05).For simple intravenous thrombolysis,bridging therapy was a protective factor for the improvement of treatment efficiency in the DTI process (OR,0.530,95%CI 0.297-0.943,P=0.031).Compared with the bridging therapy,the direct endovascular therapy was a protective factor for DTP treatment (OR,0.427,95%CI 0.202-0.901,P=0.025).The remaining independent variables were not associated with the occurrence of DTN and DTP delay (all P>0.05).(4) During the three years,the delay of intravenous thrombolysis was mainly due to objective reasons.The constituent ratio of other reasons caused delay of intravenous thrombolysis was decreased year by year.There was no other reasons for delay in the third year).There was no significant difference in the constituent ratio of the delay reasons in endovascular treatment (x2=3.622,P=0.164).Conclusion Under the existing process and resource allocation,setting the DTN target time and implementing continuous quality improvement are conducive to the effective implementation of brain CT scan and continuous optimization of intravenous thrombolysis in the processes in AIS patients with the first diagnosis.