The NMGCF-19 strain is an E.coli strain isolated and identified in our laboratory from lambs manifesting severe diarrhea and meningitis.Previous analysis of the genome sequence of NMGCF-19 strain showed that the outer membrane protein A(ompA)gene was a potential viru-lent gene.In order to determine whether the ompA gene is associated with the pathogenicity of NMGCF-19 strain and the underlying mechanism,the NMGCF-19 strain with ompA knockout(NMGCF-19△ompA)was generated in this study using CRISPR/Cas9 technology and used to de-termine the role of ompA gene in mediating the encephalitis by NMGCF-19 infection and the un-derlying mechanism using the mouse model system.The results showed that the neuronal cell nec-rosis in the hippocampus in mice infected by NMGCF-19△ompA was significantly reduced and was not focal compared with that of mice infected with the wild-type NMGCF-19 strain.The number of bacteria in brain of mice infected by NMGCF-19 △ompA was significantly reduced in comparison to that of mice infected by NMGCF-19.Simultaneously,the mRNA and protein expression levels of the tight junction proteins ZO-1 and Occludin were both increased in mice infected by NMGCF-19 △ompA strain compared with the mice infected by NMGCF-19 strain.These results suggest that the ompA gene is a virulent gene and plays an important role in the invasion of the blood-brain barrier by NMGCF-19 strain in mice.

Objective:To explore the characteristics of 24-hour movement in preschoolers and its relationship with executive function.Methods:The study subjects consisted of 555 preschoolers in Zhuhai City. An objective measurement approach was employed, physical activity level was assessed by utilizing accelerometers and executive function was evaluated by computerized behavioral tests.The characteristics of 24-hour movement in children were described.Data were analyzed using SPSS 26.0 software, independent samples t-test was used to compare differences in 24-hour activity behaviors across genders, and multivariate linear regression was used to explore the effect of preschoolers' achievement of activity behaviors on executive functioning. Results:According to the physical activity guideline for Chinese preschoolers aged 3-6 years recommendations for children's daily moderate to vigorous-intensity physical activity, sedentary screen time, and sleep duration, among the study participants, the compliance rate of moderate to vigorous-intensity physical activity (≥60 min/d) was 33.9 %(188/555), sedentary screen time (≤1 h/d) was 43.4%(241/555), and sleep duration (10-13 h/d) was 83.4 %(463/555). Only 13.3 %(74/555) of the participants met the recommended guidelines for all three behaviors. The correct score of the working memory test was (60.6±20.3) and the reaction time of the inhibitory control test was (695.1±135.8) ms. In male preschoolers, moderate to vigorous-intensity physical activity (≥60 min/d) positively predicted working memory( β=0.121, P=0.037), while sedentary screen time (≤1 h/d) negatively predicted inhibitory control ( β=-0.122, P=0.036). Conclusions:The daily compliance rate of moderate to vigorous-intensity physical activity and sedentary screen time is low among preschooler. The former is positively correlated with working memory in preschool boys, while the latter is negatively correlated with inhibitory control.

AIM:To investigate the effect of doublecortin-like kinase 1(DCLK1)on the biological properties of gastric cancer stem cells,and to explore its possible mechanism.METHODS:Serum-free suspension culture of gastric cancer stem cells and targeted inhibition of DCLK1 activity in gastric cancer stem cells with DCLK1 inhibitor DCLK1-IN-1 were performed.The expression levels of DCLK1,stemness-related proteins(SOX2 and OCT4),proliferation-related pro-teins(cyclin D1 and c-MYC),drug resistance-related proteins(ABCG2 and TOP2A),epithelial-mesenchymal transition-related proteins(E-cadherin,vimentin and Snail),and PI3K/AKT/mTOR signaling pathway-related proteins in gastric cancer stem cells were examined by Western blot.The effects of DCLK1 on viability and drug resistance of gastric cancer stem cells were determined by CCK-8 assay,and the effects of DCLK1 on self-renewal of gastric cancer stem cells were de-termined by methylcellulose spheroid-forming assay.Wound-healing and Transwell assays were performed to assess the ef-fect of DCLK1 on the migration and invasion of gastric cancer stem cells.RESULTS:The expression levels of DCLK1 and stemness-related proteins SOX2 and OCT4 in gastric cancer stem cells were significantly higher than those in parental cells(P＜0.01).The proliferation,drug resistance,migration and invasion of gastric cancer stem cells in DCLK1 inhibi-tion group were significantly lower than those in Sphere cell group(P＜0.01).The expression levels of proliferation-related proteins(c-MYC and cyclin D1)and drug resistance-related proteins(TOP2A and ABCG2)were down-regulated,the ex-pression of epithelial marker E-cadherin was up-regulated,the expression of mesenchymal markers vimentin and Snail was down-regulated,and the expression levels of PI3K/AKT/mTOR signaling pathway-related proteins and their phosphoryla-tion levels were reduced in DCLK1 inhibition group(P＜0.05).CONCLUSION:DCLK1 is highly expressed in gastric cancer stem cells,which may be involved in the proliferation,drug resistance and invasion of gastric cancer stem cells by regulating PI3K/AKT/mTOR signaling pathway.It suggests that DCLK1 can be used as a potential target for gastric cancer stem cells.

With the advances in understanding the relationships among biomaterials, the immune system and the skeletal system, the host responses elicited by implanted biomaterials can be balanced by properly designing material characteristics from the perspective of osteoimmunology. The immunoregulatory properties of bone tissue engineering scaffolds provide advantages for inducing macrophages from the proinflammatory M1 phenotype to the anti-inflammatory M2 phenotype and promoting osseointegration. Hydrogels are increasingly a focus in bone tissue engineering, and the immune response can be affected by different compositions of hydrogels, such as the sources, concentration, molecular weight, coupling with fibronectin, and the addition of cross-linking agents. Different physicochemical properties of modified hydrogels can trigger different host immune responses, modified by using soft photolithography to fabricate micropatterned hydrogels, adding enzyme-sensitive sequences, ester bonds and dynamic covalent chemistry to prevent rapid or slow degradation of the hydrogels, and adding porogens and 3D printing to modify the hydrogels with macroporous interconnective pore structures, soft and injectable hydrogels, etc. These optimized hydrogels can reduce proinflammatory factors, promote M2 macrophage polarization, and minimize foreign body reactions to evoke bone regeneration. However, the mechanism underlying the bone immune response is still poorly understood, and further study of the effects of hydrogels with different physicochemical properties on immune regulation is needed.

Qinghai University lies in western China. There are some gaps between western and eastern universities at student level, education fund, experiment condition and technique supply for graduate students. To improve educational quality for graduate students, Department of Pathophysiology of Qinghai University focused on teaching methods, evaluation approaches and capacity of teachers and constructed a course named Clinical Pathophysiology for scientific research graduate students. The course adopted a series of measures such as combining lectures with students' self-learning (PBL, paper discussion, lecture for students), introducing experts from other schools to give lectures, establishing formative evaluation, optimizing teacher's group and so on, which have gained some achievements and improved the training quality for graduate students.

Objective: To investigate the role of peroxisome proliferator-activated-receptor-γ coactivator-1α (PGC-1α) in carotid atherosclerosis due to human cytomegalovirus (HCMV) infection. Methods: Thirty-three samples of carotid arterial sclerosis plaques after carotid endarterectomy (CEA) were collected in the experiment group, and 12 pieces of normal intracranial arteries were collected in the control group. The plaques of carotid artery were studied using in situ hybridization (ISH) and immunohistochemistry (IHC) for the role of PGC-1α in HCMV related atherosclerosis. HCMV immediate early (IE) gene of two groups was detected using ISH, and the expression of PGC-1α using IHC. Results: HCMV IE gene was positive 72.7% in the atherosclerotic plague group, while 16.6% in the control group, there was significant difference between the two groups (P<0.05). As to PGC-1α expression, in the atherosclerosis group, 8/33(24.2%) were strongly positive (+ + + ), 9/33(27.2%) were positive (+ + ), 6/33(18.2%) were weakly positive (+ ), and 10/33(30.3%) were negative (-). While in the control group, 1/12(8.3%) was strongly positive (+ + + ), 2/12(16.7%) were positive (+ + ), and 1/12 (8.3%) was weakly positive (+ ), and 8/12 (66.7%) were negative (-). The differences between the two groups were statistically significant (P<0.05). Conclusions HCMV immediate early gene and PGC-1α were both related to athersclerotic plaque formation, suggesting that PGC-1α may play an important role in as the plaque formation due to HCMV infection.

Objective: To investigate the role of lectin-like Ox-LDL receptor-1( LOX-1) in the activation and oxidative stress of cultured human umbilical vein endothelial cell(HUVEC) after human cytomegalovirus(HCMV) infection. Methods: HUVEC were divided into four groups: HCMV, Control, Carrageenan, and HCMV+ Carrageenan. After HCMV AD169 infection, the supernatant of the culture was extracted, and cells were lysed. The levels of LOX-1 mRNA, intercellular adhesion molecule-1(ICAM-1) mRNA and vascular cellular adhesion molecule-1(VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxidum(NO) of the supernatant was detected by nitrate reductase method accordingly. Results: 24 h after infection, the mRNA expression of LOX-1, ICAM-1 and VCAM-1 in HUVEC of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly. The mRNA expression of LOX-1, ICAM-1 and VCAM-1 and the quantity of NO decreased after adding the LOX-1 inhibitor carrageenan. There was significant difference between groups(P<0.05). Conclusions HCMV may increase the mRNA expression of ICAM-1 and VCAM-1 and quantity of NO by upregulating the mRNA expresion of LOX-1, which may contribute to the formation of a therosclerosis(AS).

This paper aimed at investigating the effects of Xinfukang Oral Liquid on mitochondrial proteomic alterations in rats with heart failure after myocardial infarction and exploring its possible mechanisms. Heart failure models were established by ligating the left anterior descending coronary artery of SD rats. Rats were randomly divided into sham group, model group, Xinfukang group. 8 weeks after drug intervention, the mitochondrial proteomic alterations of myocardial tissue were detected by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) . Furthermore, expression levels of part of the differently expressed proteins were verified by western blot. Compared with model group, 20 differentially expressed protein spots were detected in Xinfukang group, 13 of which showed increased protein expression and 7 decreased; 13 differentially expressed protein spots were successfully identified by MALDI-TOF-MS. Bioinformatics analysis showed that these differential proteins were mainly associated with energy metabolism, stress reaction, oxidative damage, cyto-skeleton, cell differentiation and proliferation. Western blot results showed that the expression of Stress-70 protein and Nucleophosmin increased and the expression of ATP-αdecreased in model group. The expression of Stress-70 protein and Nucleophosmin decreased and the expression of ATP-αincreased in Xinfukang group, which shows the same results in proteomics. Xinfukang Oral Liquid can partly adjust proteomic alterations of myocardial mitochondrial in HF rats, and its intervention mechanism may involve improving energy metabolism, reliving stress reaction and oxidative damage, as well as regulating cell differentiation and proliferation. The results of comparative proteomic analysis performed by using2-DE and MALDI-TOF-MS are acurate, stable and reliable.

The core symptoms of Parkinson's disease include motor symptoms and non-motor symptoms. For the motor symptoms, the rehabilitation includes the basic training of joints, gait training, the training of core muscle group and posture balance, the training of oral, facial, neck muscle and pharyngeal reflex, the training of lip movement, vocalization, sound volume, rhythm and speed, and the training of deep breathing exercises and abdominal pressure. For the non-motor symptoms, the rehabilitation includes the training of memory, attention, problem solving ability, activities of daily living and others, psychotherapy, family support, social participation, and other comprehensive treatments combined with medicine.

In order to adapt to the demand of culturing high quality talents and to strengthen student's comprehensive quality,pathophysiology department of medical college of Qinghai University carried on the reform on the pathophysiological experiment examination system.After researches and practices in recent years,we established a set of relatively sound examination system including experiment operating and experiment report in normal study,report for designable experiment and experiment skill and theory exam.Investigation results demonstrated that examination system after reform functioned well in training the students' operation ability,observation ability and ability to analyze and solve questions.