BACKGROUND:Cellular senescence is closely related to the development and progression of osteoarthritis,but the specific targets and regulatory mechanisms are not yet clear. OBJECTIVE:To mine key genes in cellular senescence-mediated osteoarthritis by integrating bioinformatics and machine learning approaches and validate them via experiments to explore the role of cellular senescence in osteoarthritis. METHODS:The osteoarthritis gene expression profiles obtained from the GEO database were intersected with cellular senescence-related genes obtained from the CellAge database and the expression of the intersected genes was extracted for differential analysis,followed by GO and KEGG analysis of the differential genes.The key osteoarthritis cellular senescence genes were then screened by protein-protein interaction network analysis and machine learning,and in vitro cellular experiments were performed.Finally,the expression of the key genes was detected by qPCR. RESULTS AND CONCLUSION:A total of 31 osteoarthritis cell senescence differential genes were identified.GO analysis showed that these genes were mainly involved in the biological processes,such as regulation of leukocyte differentiation,monocyte differentiation,regulation of T cell differentiation and exerted roles in DNA transcription factor binding,histone deacetylase binding,chromatin DNA binding,and chemokine binding.KEGG analysis showed that osteoarthritis cell senescence differential genes were mainly activated in the JAK/STAT signaling pathway,PI3K/Akt signaling pathway and FoxO signaling pathway.MYC,a key gene for osteoarthritis cellular senescence,was identified by protein-protein interaction network topology analysis and machine learning methods.The results of the in vitro cellular assay showed that the mRNA expression of MYC was significantly lower in the experimental group(osteoarthritis group)than the control group(normal group)(P＜0.05).To conclude,MYC can be a key gene in the senescence of osteoarthritic cells and may be a new target in the prevention and treatment of osteoarthritis by mediating immune response,inflammatory response and transcriptional regulation.

Objective To observe the protective effect of Zhilong Huoxue Tongyu Capsule(Zhilong Capsule)on myocardial ischemia reperfusion injury(MIRI)in mice,and explore its regulatory mechanism using metabolomics.Methods Using a random number table method,30 C57BL/6J mice were randomly divided into the following three groups:sham operation group,model group,and Zhilong Capsule group(6.24 g/kg),with 10 mice in each group.In mice in the model group and the Zhilong Capsule group,a mouse MIRI model was established by ligating the left anterior descending branch,while mice in the sham operation group underwent threading without ligation.The Zhilong Capsule group began modeling one week after pre-administration and continued to receive intragastric administration for two weeks after modeling once daily.The cardiac function,including the left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS),was assessed by color echocardiography.The myocardial fibrosis and apoptosis were observed by Masson staining and TUNEL staining,respectively.Enzyme-linked immunosorbent assay was used to measure the serum contents of lactate dehydrogenase(LDH)and brain natriuretic peptide(BNP).Liquid chromatography-mass spectrometry combined with multivariate statistical method was performed for serum metabolite detection and identification analysis.Results Compared with the model group,the mice in the Zhilong Capsule group exhibited an increase in LVEF and LVFS,a reduction in cardiac tissue structure disorder,a decrease in myocardial fibrosis,a decrease in cell apoptosis rate,and a decrease in serum LDH and BNP contents(P<0.05).Metabolomics result showed that intervention with Zhilong Capsule significantly regulated 30 differential metabolites related to MIRI.Important metabolic pathways involved 20 pathways related to tyrosine metabolism,arginine and proline metabolism,and vitamin digestion and absorption.Conclusion Zhilong Capsule has a protective effect on MIRI,and it may achieve this effect by regulating pathways related to tyrosine metabolism,arginine and proline metabolism,and vitamin digestion and absorption.

Objective:To analyze the activation of primary somatosensory cortex(S1)neurons in post-traumatic stress disorder(PTSD)mice after tactile stimulation of whiskers and the changes of S1 cortical neurons in PTSD mice.Methods:Using the neuron cytoskeleton-associated protein(Arc)labeling strategy and immunofluorescence staining technique,the Arc of S1 cortical neurons in PTSD mice and control mice after whisker stimulation was marked and ob-served.By analyzing the difference in the spatial expression position of Arc labeled neurons and the number of positive cells,the activation level of S1 cortical neurons in the two groups was compared and analyzed.The pyramidal neurons of S1 cortex were labeled by sparse virus labeling method,and the number of dendrites and the morphology and number distribution of dendritic spines were compared between the two groups.Results:After whisker stimulation,it was found that Arc positive neurons were distributed from shallow layer to deep layer of S1,and more densely distributed in layersⅡ/Ⅲ and V.Compared with the control group,the number of positive neurons in different layers of the PTSD group was significantly increased.The results of cell morphology and structure analysis showed that,compared with the control group,the density of dendritic spines in layer Ⅱ/Ⅲ of mice with PTSD increased,and the number of mushroom dendrit-ic spines increased,while the number of filamentous pseudopod dendritic spines decreased.The number of dendritic spines of Si V layer mushroom type and slender type was higher,but the total number of dendritic spines was not sig-nificantly different.Conclusion:After whisker stimulation in PTSD mice,S1 neurons were over-activated,and the structure and morphology of neurons changed significantly.

Objective:To systematically summarize the psychological experience of cancer patients caregivers, and comprehensively understand their inner needs, so as to provide basis for better clinical medical work.Methods:CNKI, Wanfang DATA, VIP, EMBASE, PubMed, BMJ Journals, CINAHL, Cochrane Library, ProQuest, and other databases were searched from their inception to May 2020, to collect qualitative studies on psychological experience of cancer patients caregivers. The quality of included studies was evaluated according to JBI Critical Appraisal Tool for Qualitative Studies in Australia. The results were integrated by integrating methods.Results:A total of 10 studies were included. 22 research results were extracted and grouped into 9 new categories according to similarities. These categories resulted in 3 synthesized findings: caregivers suffered from physical and psychological sufferings, and were tortured with mainly negative emotions and mixed positive emotions; after self adjustment, the role of caregiver grew; their thirst for information and support.Conclusion:Healthcare workers should pay attention to psychological changes and information needs of cancer patients caregivers, provide necessary support to help them adapt their roles, and improve their quality of life.

Objective: To understand the consciousness of the cancer early treatment and its demographic and socioeconomic factors. Methods: A cross-sectional survey was conducted in 16 provinces covered by the Cancer Screening Program in Urban China (CanSPUC) from 2015 to 2017. A total of 32 257 local residents aged ≥18 years old who could understand the investigation procedure were included in the study by using the cluster sampling method and convenient sampling method. All local residents were categorized into four groups, which contained 15 524 community residents, 8 016 cancer risk assessment/screening population, 2 289 cancer patients and 6 428 occupational population, respectively. The questionnaire collected personal information, the consciousness of the cancer early treatment and relevant factors. The Chi square test was used to compare the difference between the consciousness of the cancer early treatment and relevant factors among the four groups. The logistic regression model was used to analyze the influencing factors related to the consciousness of the cancer early treatment. Results: With the assumption of being diagnosed as precancer or cancer, 89.97% of community residents, 91.84% of cancer risk assessment/screening population, 93.00% of cancer patients and 91.52% of occupational population would accept active treatments (P<0.001). If the immediate family members were diagnosed as precancer or cancer, people who would encourage their family members to receive early treatment in the four groups accounted for 91.96%, 91.94%, 92.44% and 91.55%, respectively (P<0.001). The company employees, annual household income with 40 000 yuan and more and other three groups had a relatively better consciousness of the cancer early treatment (P<0.05). Male, widowed, unemployed and from the central and western regions had a relatively worse consciousness of the cancer early treatment (P<0.05). Conclusion Residents in urban China participants had a good consciousness of the cancer early treatment. The marital status, occupation, annual household income and residential regions were major factors related to the consciousness of the cancer early treatment.

Objective: To understand the prevalence of carotid plaque (CP) in population at high-risk for cardiovascular disease (CVD) in Jiangsu province and identify related influencing factors. Methods: Based on the China Patient-centered Evaluative Assessment of Cardiac Events Million Persons Project from 2015 to 2016, a total of 11 392 persons at high-risk for CVD were selected from six project areas in Jiangsu province for the questionnaire survey, physical measurement, laboratory test and bilateral ultrasound examination of carotid arteries. The prevalence of CP and influencing factors of abnormal carotid arteries, CP and plaque burden (CP≥2) were analyzed. Results: Among the persons surveyed, 4 821 (42.3%) were males. The age of the persons surveyed was (59.4±8.9) years. There were 5 971 abnormal carotid arteries cases (52.4%), including 1 782 carotid intima-media thickness thickening cases (15.6%), 3 811 CP cases (33.5%) and 378 carotid stenosis cases (3.3%). Older age (OR=2.253, 95%CI: 2.127-2.386), urban residence (OR=2.622, 95%CI: 2.375-2.895), hypertension (OR=1.439, 95%CI: 1.195-1.732), smoking (OR=1.441, 95%CI: 1.259- 1.650), pulse pressure difference (OR=1.270, 95%CI: 1.198-1.347), fasting plasma glucose (FPG) (OR=1.109, 95%CI: 1.059-1.161) and LDL-C/HDL-C (OR=1.225, 95%CI: 1.164-1.288) were possible risk factors of CP in population at high risk for CVD. Being women (OR=0.558, 95%CI: 0.494-0.630), high BMI (OR=0.948, 95%CI: 0.904-0.994), higher levels of education (OR=0.708, 95%CI: 0.531-0.945), and higher annual household income (OR=0.773, 95%CI: 0.669-0.894) were the possible protective factors. Conclusions Over half of the population at high-risk for CVD in Jiangsu showed abnormal carotid arteries. High blood pressure, high blood glucose, high blood lipids and smoking were the main factors that could be changed.

Objective To profile 16S rRNA methylase genes and the flanking sequences in extensively drug resistant (XDR) Pseudomonas aeruginosa. Methods A total of 59 strains of XDR P. aeruginosa were collected. MICs of antimicrobial agents against these strains were determined by agar dilution method. The genes encoding 16S rRNA methylase (armA, rmtA, rmtB, rmtC, rmtD, rmtE, and npmA) were analyzed by PCR. The homology of strains was studied by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR). The structure of armA and rmtB flanking regions were characterized. Results The overall prevalence of armA or rmtB genes was 62.7％ (37/59) in XDR P. aeruginosa isolates, specifically armA positive in 17 strains and rmtB positive in 22 strains. The rmtA, rmtC, rmtD, or npmA gene was not identified in any strain. ERICPCR generated 19 types (or clones) from the 59 strains. The 17 armA-positive strains were distributed in 3 clones, and 88.2％ of the armA-positive strains (15) belonged to clone D. The rmtB gene was dispersed in 9 clones. Flanking sequence analysis demonstrated that armA gene was located in a mobile element carrying multiple transposases. The sequence of the mobile element showed 99％ similarity with the sequence of armA-positive plasmid carried by E. coli and A. baumannii. The rmtB gene was also located in a mobile element containing multiple transposases. The sequence of the mobile element was highly consistent with that of rmtB-positive plasmid carried by E. coli and K. pneumoniae. Conclusions The genes encoding 16S rRNA methylase are prevalent in XDR P. aeruginosa strains. All the genes were identified in high-level gentamicin-resistant strains. Clonal dissemination may explain the spread of armA among P. aeruginosa isolates.

Objective To investigate the resistance profile of Klebsiella pneumoniae isolates in Huashan Hospital, Fudan University. Methods The MICs of fluoroquinolones were determined by agar dilution method against 112 clinical strains of K. pneumoniae. Multilocus sequence typing (MLST) were applied to 48 K. pneumoniae strains. The characteristic sequence type (ST) associated with antibiotic resistance was identified by PCR. Results Lower percentage (＜40％) of K. pneumoniae strains were susceptible to fluoroquinolones. Majority (86.2％) of ciprofloxacin non-susceptible K. pneumoniae strains belonged to CC1 (ST11), ST494 or CC4 (ST15 and ST655), indicating the potential of clonal dissemination. ST494 (18.8％) was the second commonest sequence type, next only to ST11. ST494 strains harbored the genes encoding beta-lactamases, oqxAB, qnrD, aac-(6')-lb-cr and armA and had a single point mutation in gyrA. Therefore, ST494 strains were highly resistant to cephalosporins, fluoroquinolones and aminoglycosides and 22％ of the strains were resistant to carbapenems. However, all the ST494 strains were susceptible to tigecycline and tetracycline. Conclusions ST11 and ST494 are the commonest STs of K. pneumoniae conferring multidrug resistance in this hospital. These STs may contribute to the high resistance rates of K. pneumoniae to fluoroquinolones. The susceptibility of ST494 strains to tigecycline and tetracycline allows us to consider the promising potential of such drugs in managing K. pneumoniae infections.

Objective To investigate the effect of different doses of ketamine on inflammatory cytokines in rabbits with severe burn at early stage and preliminarily approach its regulatory action on early stage of inflammatory reaction due to stress of trauma.Methods Forty healthy male New Zealand rabbits were randomly divided into four groups in accord with the random number table method: normal control group, scald model group, ketamine analgesia group and ketamine anesthesia group. Before scald, pentobarbital sodium was used for anesthesia, afterwards catheters were inserted into internal jugular vein and internal carotid artery respectively ready for use, and 24 hours later, Ⅲ degree scald at the animal back and buttocks occupying 30% total body surface area (TBSA) was performed as the scald model for all the rabbits except those in normal control group. In ketamine analgesia group, after scald for 0.5 hour, 0.5 mg/kg ketamine intravenous injection was given to the rabbits as the loading dosage and then persistent intravenous pump infusion of 9μg·kg-1·min-1 ketamine was applied for all together 24 hours. In ketamine anesthesia group, after scald for 0.5 hour, 1.5 mg/kg ketamine intravenous injection was given to the rabbits, and then persistent intravenous pump infusion of 45μg·kg-1·min-1 ketamine was applied for 4 hours to maintain systemic anesthesia. In normal control and scald model groups, only intravenous infusion of equal amount of normal saline was given to the rabbits. The amount of intravenous transfusion in each group and the total dosages of ketamine used in ketamine analgesia group and ketamine anesthesia group were recorded. Before scald and 0.5, 6, 12, 24 hours after scald, arterial blood gas analyses were made, and the levels of serum interleukins (IL-1, IL-6) and tumor necrosis factor-α (TNF-α) were determined.Results Although the indexes of blood gas analysis were changed in the four groups, they were all in the normal range, showing that the respiratory function was in the normal range and indirectly reflecting that the circulatory function was also in the normal range, thus the effects on cytokines by factors of respiratory and circulatory functions were ruled out. The levels of IL-1, IL-6 and TNF-α before scald showed no statistically significant differencesamong the four groups (allP > 0.05). From 0.5 hour after scald, the levels of IL-1, IL-6 and TNF-α were markedly higher in model group than those of normal control group [IL-1 (ng/L): 30.27±0.93 vs. 13.79±1.11, IL-6 (ng/L): 47.22±1.49 vs. 46.31±4.12, TNF-α (ng/L): 243.39±20.85 vs. 190.95±14.97, allP < 0.05], and the situation continued until 24 hours after scald; the levels of IL-1, IL-6 and TNF-α from 6 hours after scald were significantly decreased in ketamine analgesia and ketamine anesthesia groups compared with those in the model group, and from 12 hours after scald, the degrees of descent in levels of the above indexes in ketamine analgesia group were more obvious than those in ketamine anesthesia group [IL-1 (ng/L): 19.28±2.51 vs. 40.12±10.31, IL-6 (ng/L): 52.10±4.23 vs. 72.20±10.11, TNF-α (ng/L): 246.03±20.74 vs. 313.71±27.34, allP < 0.05].Conclusion The low-dose ketamine analgesia and ketamine anesthesia have certain degree of inhibitory effect on the expression and release of inflammatory cytokines at the early stage in rabbits with severe burn, the effect of long-term low-dose ketamine analgesia being more significant.

Objective To investigate the possibility of chloroquine radiosensitization of esophageal cancer cell line TE-1 and its further mechanism.Methods Effect of chloroquine on cell viability of TE-1 cells was determined by MTT method.Expression of LC3,Beclin-1 and formation of acidic vesicular organelles (AVOs) were determined by Western blot,and fluorescence staining with Lyso-Tracker Red DND-99,respectively.Clonogenic survival of TE-1 cells was examined by clonogenic forming assay.Results Chloroquine showed dose-dependent inhibition of TE-1 cell growth,and its values of IC50 and IC10 were (72.33 ± 5.28) and (15.42 ± 3.33) μmol/L,respectively.The expression of Beclin-1 and LC3-Ⅱ/Ⅰ markedly increased in irradiated TE-1 cells.The addition of chloroquine with IC10concentration significantly reduced the fluorescence and intensity of AVOs accumulation in the cytoplasm of TE-1 cells.Clonogenic survival fraction decreased obviously,in TE-1 cells with addition of chloroquine after radiation and the value of SERD0 was 1.439.Conclusions Chloroquine could radiosensitize esophageal cancer cells by blocking autophagy-lysosomal pathway and be used as a potential radiosensitizing strategy.