OBJECTIVE To investigate the dynamic changes in urine metabolic profiles in rats induced by D-galactose (D-Gal),and to study the correlations between the differential metabolites and behavior indicators using the proton nuclear magnetic resonance (1H-NMR)-based metabonomics.METHODS Subcutaneous injection of D-Gal 100 mg· kg-1 for 10 weeks was adopted in the model group.The sample of urine was collected at day 0 (dO),d14,d28,d42,d56 and d70.NMR metabonomics technique was used for acquisition of data,which was analyzed by multivariate statistical analysis.The ability of learning and memory were measured by Morris water maze test from d70.After the behavioral test,the rats were sacrificed and the hippocampus was observed by hematoxylin-eosin staining.RESULTS Principal component analysis (PCA) results revealed that there was considerable difference between the model group and the normal control group at d70.According to the varible importance plot (VIP) calculation and S-plot scores,a total of 12 metabolites were screened and identified as potential biomarkers at d70.The differences of metabolites and Morris water maze test were subjected to correlation analysis,and the results showed that the levels of choline,lactate and dimethylglycine in the model group were significantly increased and negatively correlated with the times of crossing the platform (r =-0.90,-0.50 and-0.52;n=10).Formate was significantly negatively correlated with the time spent in the target area (r =-0.51,n=10),but choline and formate were significantly positively correlated with the escape latency (r =0.72 and 0.53;n=10).However,the levels of creatine and taurine decreased in the model group,which was significantly positively correlated to the times of acrossing platforms (r =0.89 and 0.71;n=10),while alanine was significantly positively correlated to the time spent in the target area(r =0.74,n=10).Taurine,alanine and creatine were significantly negatively correlated with the escape latency (r =-0.66,-0.50 and-0.85;n=10).The correlations between the differential metabolites and the behavioral indicators were further proved.CONCLUSION The metabolic profile changes in urine from D-Gal induced aging model rats are significantly correlated with impairement of ability in learning and memory.1H-NMR metabonomics in urinary metabolic profile changes may be used as an evaluation index in the D-Gal induced aging rats model.

Objective:To analyze the differentally expressed long non-coding RNA (lncRNA) among mice of different ages and explore the mechanism of kidney aging.Methods:Male C57BL/6 mice aged 3-month-old ( n=5), 12-month-old ( n=5) and 24-month-old ( n=5) (each weighting about 25 g) were randomly selected. PAS staining, Masson staining and senescence associated β-galactosidase (SA-β-gal) staining were used to detect the pathology and cell senescence of mice kidney. High throughput sequencing was performed to detect the differentially expressed lncRNA and their fragments per kilobase million. Real-time quantitative PCR was used to verify the differentially expressed lncRNA. Competitive endogenous RNA (ceRNA) network, which consisted of lncRNA, miRNA and mRNA was built. GO and KEGG enrichment analysis method were used to predict the biological function of differentially expressed lncRNA. Results:PAS staining and Masson staining showed the development of kidney fibrosis, and SA-β-gal staining positive region was increased significantly as age increased. There were 938 known lncRNA and 542 novel lncRNA differentially expressed among different ages' mouse kidney. Compared with 3-month-old mice, 33 lncRNA were up-regulated and 43 lncRNA were down-regulated in 12-month-old mice. Compared with 3-month-old mice, 130 lncRNA were up-regulated and 91 lncRNA were down-regulated in 24-month-old mice. Compared with 12-month-old mice, 36 lncRNA were up-regulated and 22 lncRNA were down-regulated in 24-month-old mice. The results of qRT-PCR about verified 10 lncRNAs with larger differential expression multiples and higer expression levels were consistent with the sequencing data. GO enrichment analysis showed that the target genes of lncRNA differentially expressed in the three groups were mostly located in the nucleus and cytoplasm, and might play a role by binding to proteins or participate in various protein phosphorylation, cell cycle, transcription, transcription regulation and other processes. KEGG enrichment analysis showed that the target genes of lncRNA differentially expressed in the three group were significantly enriched in Rap1 signaling pathway, FOXO signaling pathway and MAPK signaling pathway, which were closely related to kidney aging.Conclusion:There are significant differences in expression of lncRNA among the kidney of different ages mice, which are involved in the occurrence of renal senescence.

Objective To assess the effects of early correction of anemia with recombinant human erythropoietin (rHuEPO) on the development and progression of left ventricular hypertrophy (LVH) in patients with mild-to-moderate chronic renal insufficiency (CRI) who are not on hemodialysis. Methods A total of 158 patients with serum creatinine from 147μmol/L to 400μmol/L were nrolled in this prospective, multicenter study. Eighty-six patients with hemoglobin (Hb)＜110g/L received rHuEPO treatment with a target Hb of ≥110g/L (Group A). Forty patients with comparable Hb concentration (＜110g/L) but did not receive rHuEPO (Group B) and 32 patients with Hb≥110g/L and without rHuEPO treatment (Group C) were served as controls. Left ventricular mass index (LVMI) was evaluated by echocardiography at baseline and every 3 months for 2 years. Results There was no difference in age, gender, etiology of renal failure, blood pressure and cardiovascular risk factors among the 3 groups. At baseline, the prevalence of LVH was 72.1% in group A,72.5% in group B and 59.4% in group C. LVMI was inversely correlated with Hb levels (r=0.70, P＜0.01). During the 2-year period, the mean LVMI decreased from 142.6±25.7g/m2 to 132.4±18.5 g/m2 in group A, while increased significantly in both group B and group C. The mean Hb concentration increased from 93.8±14.6g/L to 111.2±10.3g/L (P＜0.05) in group A, but tended to decrease in group B and group C. There was no significant change of the mean blood pressure, number of anti-hypertensive drugs and serum creatinine concentrations in all 3 groups. However, patients' serum creatinine doubled more often in group B and group C than in group A.Conclusions LVH was common in predialysis CRI patients and was associated with the severity of anemia. Early intervention with rHuEPO may reverse LVH in these patients.

Depression is a complex mental disease with polygenic inheritance and a high incidence.Our understanding of the clinical manifestations and pathogenesis of depression has recently improved.Continuous progress in gene-editing technologies has increased the construction efficiency and reduced the cost of gene-knockout animals,leading to their increasing use in the fields of basic research and drug development for depression and providing a powerful tool for revealing the pathogenesis of depression.In this review,we summarize recent progress in understanding the roles and mechanisms of candidate genes in depression using knockout model mice.

Objective:To explore the relationship between triglyceride-glucose (TyG) index with plaque components, plaque burden and characteristics of vulnerable plaque using coronary plaque analysis based on coronary artery computed tomography (CCTA).Methods:A total of 498 patients(male 296, female 202), the age ranged from 33 to 87 (63±9) years who underwent CCTA from January 2020 to September in Shandong Provincial Hospital Affiliated to Shandong First Medical University were included. The enrolled patients were divided into three groups according to the tertiles of TyG index: T 1 group (the lowest one-third), T 2 group (middle one-third) and T 3 group (the highest one-third). The plaque burden, volume and ratio of calcified, lipid and fibrous components of plaques as well as the incidence of vulnerable plaques were measured based on CCTA images. Chi-square test, ANOVA and Kruskal-Wallis test were used to compare whether the differences of indexes among the three groups were statistically significant. Multiple stepwise regression was used to analyze the influencing factors of coronary atherosclerotic plaque burden and multiple logistic regression was used to analyze the risk factors of CT high-risk plaque. Finally, ROC curve was used to evaluate the value of TyG index in the diagnosis of CT high-risk plaque, and the best diagnostic threshold of TyG index was determined. Results:The plaque burden, non-calcified plaque volume and ratio had positive relationship with TyG index ( P<0.001).TyG index was significantly correlated with plaque burden according to multiple stepwise regression analysis (regression coefficient 7.267, P<0.001). The results of CT vulnerable characteristics of plaques showed that positive remodeling, low-attenuation plaque sign and the incidence of vulnerable plaque increased with TyG index ( P<0.05). Multivariate Logistic regression analysis showed that TyG index was an independent risk factor for CT vulnerable plaque(OR=2.324,95 %CI 1.533-3.523, P<0.001). The cut-off value of TyG index that can predict vulnerable plaque was 8.43(sensitivity 77.24%, specificity 45.60%, AUC 0.645, P<0.001). Conclusions:With the increase of TyG index, the burden of coronary atherosclerosis plaque and the incidence of CT vulnerable plaque increased. TyG index is expected to be a simple and effective predictor of cardiovascular disease and adverse cardiovascular events.

Objective:To investigate the value of CT features in predicting the invasion and degree of invasiveness of lung pure ground-glass nodules (pGGN) in the new histological classification in 2021.Methods:A total of 281 patients (304 lesions) with pGGN confirmed by surgical pathology from December 2018 to January 2021 in Shandong Provincial Hospital Affiliated to Shandong First Medical University were retrospectively analyzed. According to the pathological types, the patients were divided into prodromal lesion group [atypical adenomatous hyperplasia (AAH) and adenocarcinoma in situ (AIS), 129 cases], minimally invasive group [minimally invasive adenocarcinoma (MIA), 116 cases] and invasive group [invasive adenocarcinoma (IAC), 59 cases]. Clinical data (age, gender, smoking history, family history of cancer), and CT parameters [shape, boundary, lobulation, burr, vacuolar sign, bronchial abnormality sign, internal vessel sign, pleural traction sign, longest diameter, shortest diameter, unenhanced CT value, contrast-enhanced CT value in arterial phase, contrast-enhanced CT values in venous phase, the degree of enhancement (ΔCT A-N, ΔCT V-N)] were recorded and measured. The ANOVA, Kruskal-Wallis H and χ 2 test were used to compare the differences among the three groups. Binary logistic regression analysis was used to evaluate the independent risk factors of nodular invasion [prodromal lesion and invasive lesion (MIA and IAC)] and the degree of nodular invasion (MIA and IAC), and receiver operating characteristic (ROC) curve analysis was performed for each parameter. Results:There were statistically significant differences in age, pGGN morphology, lobulation, vacuolar sign, bronchial abnormality sign, internal vascular sign, pleural traction sign, longest diameter, shortest diameter, unenhanced CT value, contrast-enhanced CT value in arterial phase, contrast-enhanced CT value in venous phase among the precursor lesion group, minimally invasive group and invasive group ( P<0.05). Binary logistic regression analysis showed that vacuole sign (OR=2.832, 95%CI 1.363-5.887, P=0.005), internal vascular sign (OR=3.021, 95%CI 1.909-4.779, P<0.001) and unenhanced CT value (OR=1.003, 95%CI 1.001-1.006, P=0.019) were independent risk factors for invasion. Lobulation (OR=5.739, 95%CI 2.735-12.042, P<0.001), internal vascular sign (OR=1.968, 95%CI 1.128-3.433, P=0.017) and unenhanced CT value (OR=1.004, 95%CI 1.001-1.008, P=0.012) were independent risk factors for the degree of invasiveness. ROC curve analysis showed that the efficiency of internal vascular sign was the highest in distinguishing precursor lesion and the invasive, the area under the curve (AUC) was 0.757, the sensitivity was 50.3%, the specificity was 89.8%. The efficiency of lobulation was the highest in distinguishing MIA and IAC (AUC=0.702), with a sensitivity of 61.0% and specificity of 79.3%. Conclusions:CT features are of certain value in predicting the invasion and degree of invasiveness of lung pGGN in the new histological classification in 2021, and internal vascular sign is more effective in predicting the invasion of lung pGGN. Lobulation can predict the degree of invasiveness of pGGN better.

Objective:To explore the relative genes that may influence kidney aging and verify the expression of clock gene Arntl in aging kidney. Methods:The differentially expressed genes between C57BL/6 male aging mice (24 months old) group and young mice (3 months old) group were identified by whole transcriptome sequencing, and the enriched biological pathways and key proteins were analyzed by bioinformatics methods. RT-qPCR and Western blotting were used to verify the mRNA and protein expression of Arntl.Results:(1) A total of 119 differentially expressed genes were screened between aging mice group and young mice group by whole transcriptome sequencing. Differentially expressed genes were mainly enriched in biological processes such as rhythmic process, circadian rhythm and circadian regulation of gene expression (all P<0.001). Protein-protein interaction analysis results showed that Nfil3, Hspa8, Arntl, Hlf, Rorc, Per3 and Npas2 and so on, were the key proteins in these differentially expressed genes. The results of RT-qPCR confirmed that the expression differences of clock genes Arntl, Nfil3, Npas2 and Per3 between aging mice group and young mice group were consistent with sequencing results (all P<0.05). (2) Compared with C57BL/6 young mice group and SAMR1 rapidly aging mice, the protein expression of Arntl in aging mice group and SAMP8 rapidly aging mice had downward trends. Conclusions:Clock genes and their circadian biological pathways may play an important role in the process of renal aging. The expression of Arntl in aging kidney has a downward trend.

Objective:To explore the mediating role of interdisciplinary cooperation ability between the level of hospice care practice and difficulty degree of hospice care for medical staff, in order to improve the quality of hospice care services for medical staff, and to provide evidence for relevant managers to formulate effective measures to reduce the implementation of hospice care difficulties.Methods:This study was a cross-sectional survey. Using convenience sampling method, the general data questionnaire, Palliative Care Difficulties Scale (PCDS), Palliative Care Self-Report Practice Scale (PCPS) and Modified Index of Interdisciplinary Collaboration (MIIC) were used to investigate 362 medical staff in all hospice care institutions of Shiyan City from July to August 2022.Results:The total score of PCDS in 362 medical staff was (40.58±13.44) points, (67.47 ± 12.50) points for PCPS and (108.36 ± 21.46) points for MIIC. There was a positive correlation between the total score of MIIC and PCDS ( r=0.500, P<0.01). The total score of PCPS was negatively correlated with the total score of MIIC ( r=-0.337, P<0.01) and the total score of PCDS ( r=-0.189, P<0.01). The interdisciplinary cooperation ability of medical staff in hospice care had a complete mediating effect between the level of practice and the degree of difficulty (Effect value=-0.190, 95% CI-0.274 to -0.126), and the mediating effect accounted for 98.5% of the total effect value. Conclusions:The practice level of hospice care can not directly affect the implementation of the difficulty degree, but by taking active measures to improve the interdisciplinary cooperation ability, so as to enhance the practice level, and then reduce the implementation of hospice care difficulty degree.

Objective:To construct the second and fourth generations of CAR-T cells targeting fibroblast activation protein (FAP) on the surface of stromal carcinoma-associated fibroblasts and compare their characteristics in vitro and in vivo. Methods:ELISA was used to detect the cytokines secreted by CAR-T cells. Cell proliferation and viability were analyzed by counting. Chemotactic ability was tested by Transwell migration assay. Distribution of T cell subsets was analyzed by flow cytometry. Cytotoxicity was assessed by luciferase bioluminescence. The safety and therapeutic effects were evaluated in a NOG mouse model of metastatic human lung cancer.Results:The expression rates of the second and fourth generations of CAR-T cells (h4BBz CAR-T and h4BBz-7.19 CAR-T) were (74.280±4.384)% and (67.220±4.013)%, respectively. The h4BBz-7.19 CAR-T cells had better in vitro proliferation and chemotactic activity than h4BBz CAR-T cells as they were able to secrete IL-7 and CCL19, while the viability of h4BBz-7.19 CAR-T cells was comparable to that of h4BBz CAR-T cells. There was no significant difference in the expression rate of h4BBz CAR or h4BBz-7.19 CAR between CD4 + T and CD8 + T cells. The proportions of both Naive cells and T memory stem cells (TSCM) in CD4 + and CD8 + T cells were higher in h4BBz-7.19 CAR-T cells than in h4BBz CAR-T cells. Moreover, h4BBz-7.19 CAR-T cells possessed stronger specific cytotoxicity on the target cancer cells than h4BBz CAR-T cells when the ratio of effectors/targets was low ( P1∶1=0.004, P2∶1=0.000 6, P5∶1<0.000 1, P10∶1=0.022, P20∶1=0.116), while the expression of PD-1 on the surface of h4BBz-7.19 CAR-T cells was lower than that on h4BBz CAR-T cells. In the NOG mouse model of metastatic human lung cancer, h4BBz-7.19 CAR-T cells could slow the tumor growth and prolong the survival time of mice without causing weight loss or pathological changes in the organs. Conclusions:The fourth-generation CAR-T cells targeting FAP were shown to have stronger proliferation, better penetration and more potent specific cytotoxicity by secreting IL-7 and CCL19 and could slow the tumor growth and prolong survival by improving tumor immunosuppressive microenvironment. This study provided reference for the clinical application of the fourth generation of CAR-T cells.

Objective To explore the correlation between childhood trauma and plasma adiponectin levels in patients with depression.Methods A total of 121 patients with depression and 39 healthy controls(control group)were enrolled.Childhood trauma questionnaire(CTQ-SF)was used to assess the experience of childhood abuse and neglect,and the patients with depression were divided into trauma group(n=53)and non-trauma group(n=68)according to the CTQ-SF score.The 17-item Hamilton depression scale-17(HAMD17)and the Hamilton anxiety scale(HAMA)were used to evaluate the severity of depression and anxiety symptoms,respectively.Plasma adiponectin levels of subjects were measured by enzyme-linked immunosorbent assay.Results The plasma adiponectin level of trauma group[3.82(2.44,4.92)μg/mL]was significantly lower than that of non-trauma group[4.64(2.98,6.43)μg/mL,P=0.01]and the control group[6.29(4.54,7.51)μg/mL,P＜0.01].The plasma adiponectin level of non-trauma group was lower than that of the control group(P＜0.01).Correlation analysis showed that plasma adiponectin level in patients with depression was negatively correlated with childhood trauma(r=-0.34,P＜0.01).Multivariate linear regression analysis showed that plasma adiponectin level was negatively correlated with childhood trauma scores in patients with depression(β=-0.05,P＜0.01).Conclusions Patients with depression who have experienced childhood trauma have lower plasma levels of adiponectin,and childhood trauma may be associated with decreased plasma adiponectin levels in patients with depression.