OBJECTIVE: Myocardial inflammation during myocardial infarction (MI) could be inhibited by regulating arachidonic acid (AA) metabolism. Recent studies demonstrated that Sini Decoction (SND) was identified to be an effective prescription for treating heart failure (HF) caused by MI. But the anti-inflammatory mechanism of SND remained unclear. The work was designed to investigate the anti-inflammatory mechanism of SND through the AA metabolism pathway in vitro and in vivo experiments. METHODS: An inflammatory injury model of H9c2 cells was established by lipopolysaccharide (LPS)-stimulated macrophage-conditioned medium (CM). The MI model was built by the ligation of left anterior descending (LAD) branch of coronary artery in rat. Meanwhile, the rats were divided into five groups: sham group, MI group, MI + Celecoxib group, MI + low-dose SND group (SND-L) and MI + high-dose SND group (SND-H). Cardiac function, histopathological changes and serum cytokines were examined four weeks later. Western blot analysis was conducted to verify the key enzymes levels in the AA metabolic pathway, including phospholipase A2 (PLA2), cyclooxygenases (COXs) and lipoxygenases (LOXs). RESULTS: These in vivo results demonstrated that SND could improve the cardiac function and pathological changes of rats with MI, and regulate the key inflammatory molecules in the AA metabolism pathway, including sPLA2, COX-1, COX-2, 5-LOX and 15-LOX. In vitro, SND could decrease the release of pro-inflammatory cytokines including TNF-α and IL-6 and inhibit cell apoptosis in CM-induced H9c2 cells. Moreover, SND could protect H9c2 cells from the damage of CM by regulating nuclear factor kappa-B (NF-κB) signal pathway and the expression of COX-2. CONCLUSION SND may be a drug candidate for anti-inflammatory treatment during MI by regulating the multiple targets in the AA metabolism pathway.

Objective:To investigate the risk factors of lower extremity deep venous thrombosis (LEDVT) in patients with sepsis during hospitalization in intensive care unit (ICU), and to construct a nomogram prediction model of LEDVT in sepsis patients in the ICU based on the critical care scores combined with inflammatory markers, and to validate its effectiveness in early prediction.Methods:726 sepsis patients admitted to the ICU of the Affiliated Hospital of Jining Medical University from January 2015 to December 2021 were retrospectively included as the training set to construct the prediction model. In addition, 213 sepsis patients admitted to the ICU of the Affiliated Hospital of Jining Medical University from January 2022 to June 2023 were retrospectively included as the validation set to verify the performance of the prediction model. Clinical data of patients were collected, such as demographic information, vital signs at the time of admission to the ICU, underlying diseases, past history, various types of scores within 24 hours of admission to the ICU, the first laboratory indexes of admission to the ICU, lower extremity venous ultrasound results, treatment, and prognostic indexes. Lasso regression analysis was used to screen the influencing factors for the occurrence of LEDVT in sepsis patients, and the results of Logistic regression analysis were synthesized to construct a nomogram model. The nomogram model was evaluated by receiver operator characteristic curve (ROC curve), calibration curve, clinical impact curve (CIC) and decision curve analysis (DCA).Results:The incidence of LEDVT after ICU admission was 21.5% (156/726) in the training set of sepsis patients and 21.6% (46/213) in the validation set of sepsis patients. The baseline data of patients in both training and validation sets were comparable. Lasso regression analysis showed that seven independent variables were screened from 67 parameters to be associated with the occurrence of LEDVT in patients with sepsis. Logistic regression analysis showed that the age [odds ratio ( OR) = 1.03, 95% confidence interval (95% CI) was 1.01 to 1.04, P < 0.001], body mass index (BMI: OR = 1.05, 95% CI was 1.01 to 1.09, P = 0.009), venous thromboembolism (VTE) score ( OR = 1.20, 95% CI was 1.11 to 1.29, P < 0.001), activated partial thromboplastin time (APTT: OR = 0.98, 95% CI was 0.97 to 0.99, P = 0.009), D-dimer ( OR = 1.03, 95% CI was 1.01 to 1.04, P < 0.001), skin or soft-tissue infection ( OR = 2.53, 95% CI was 1.29 to 4.98, P = 0.007), and femoral venous cannulation ( OR = 3.72, 95% CI was 2.50 to 5.54, P < 0.001) were the independent influences on the occurrence of LEDVT in patients with sepsis. The nomogram model was constructed by combining the above variables, and the ROC curve analysis showed that the area under the curve (AUC) of the nomogram model for predicting the occurrence of LEDVT in patients with sepsis was 0.793 (95% CI was 0.746 to 0.841), and the AUC in the validation set was 0.844 (95% CI was 0.786 to 0.901). The calibration curve showed that its predicted probability was in good agreement with the actual probabilities were in good agreement, and both CIC and DCA curves suggested a favorable net clinical benefit. Conclusion:The nomogram model based on the critical illness scores combined with inflammatory markers can be used for early prediction of LEDVT in ICU sepsis patients, which helps clinicians to identify the risk factors for LEDVT in sepsis patients earlier, so as to achieve early treatment.

Objective:To investigate the effect of B7-H3 gene on the biological function of fibr-oblastlike synoviocytes (FLS) in osteoarthritis (OA).Methods:Synovial tissue of five cases of OA and synovial tissue of 4 normal knee were obtained, and the primary cell lines were isolated and cultured. The expression of B7-H3 in OA synovial tissue and primary OA-FLS were studied by immunohi-stochemistry, real time-poly merase chain reaction (PCR) and FACS. According to sites 996 and 1041 of B7-H3, corresponding siRNA was designed and the expression of B7-H3 in FLS was silenced and down-regulated. The inhibition of B7-H3 and its protein in target cells was determined by Western blot and FACS. The migration and invasion ability of B7-H3 in target cells were analyzed by scratch assay and Transwell assay. CCK8 assay was used to detect cell proliferation ability, and CBA assay was used to detect cytokines and chemokines in cell culture supernatant. GraphPad Prism 8.0 software was used to analyze the experimental data. The normal distribution data was expressed as mean±standard deviation ( Mean± SD). The comparison between data was performed by T test, and P<0.05 was considered statistically significant. Results:The abnormally high expression of B7-H3 in fibro-blast-like synoviocytes of OA was detected. Compared with siNC, si996 and si1041 inhibited the expression of B7-H3 in OA-FLS. In the Transwell migration experiment, the mean cells number of random view in the siNC group, the si996 group, and the si1041 group indicating decreased migration ability of OA-FLS [siNC vs si996 (100.3±3.7) /view vs (48.7±1.2) /view, t=13.24, P<0.001; siNC vs si1041 (100.3±3.7) /view vs (59.7±1.9) /view, t=9.80, P<0.001). In the Transwell invasion experiment, the mean cells number of random view in the siNC group, in the si996 group, and in the si1041 group indicating decreased invasion ability of OA-FLS [siNC vs si996 (127.3±5.6) /view vs (39.7±3.3) /view, t=13.49, P<0.001; siNC vs si1041 (127.3±5.6) /view vs (57.3±1.9) /view, t=11.85, P<0.001]. The secretion of IL-6 [siNC vs si996 (248±21) pg/ml vs (111±12) pg/ml, t=24.08, P=0.002; siNC vs si1041 (248±21) pg/ml vs (46±5) pg/ml, t=13.21, P=0.006], IL-8 [siNC vs si996 (118.1±15.6) pg/ml vs (47.1±5.4) pg/ml, t=6.68, P=0.022; siNC vs si1041 (118.1±15.6) pg/ml vs (10.0±1.3) pg/ml, t=13.08, P=0.006], CXCL8 [siNC vs si996 (178.8±6.4) ng/ml vs (83.2±2.7) ng/ml, t=13.77, P=0.005; siNC vs si1041 (178.8±6.4) ng/ml vs (93.5±2.8) ng/ml, t=12.23, P=0.007] and CCL2 [siNC vs si996 [(184.1±5.1) ng/ml vs (109.4±5.9) ng/ml, t=9.57, P=0.011; siNC vs si1041 (184.1±5.1) ng/ml vs (97.1±1.5) ng/ml, t=16.39, P=0.004] was decreased . Conclusion:B7-H3 may regulate the migration, invasion, cytokine secretion and other biological functions of OA-FLS, providing clues for further study of B7-H3's involvement in the pathogenesis of OA.

To construct a eukaryotic expression plasmid containing the luciferase reporter gene (Fluc) to quickly detect apoptosis. Four amino acids, Asp-Glu-Val-Asp (DEVD), the recognize motif of Caspase-3, were introduced into the middle of the Fluc-C and N fragment. Meanwhile, four amino acids, Asp-Glu-Val-Gly (DEVG), were selected as a negative control. Subsequently, the recombinant gene was cloned into the N and C terminal end of the split intein, and named as pFluc-DEVD and pFluc-DEVG. Then the plasmids were transfected into cells and renilla luciferase was co-transfected in each sample as an internal control for transfection efficiency. Then the apoptosis level was detected by the double luciferase reporter gene and the Western blotting analysis. The results showed that when apoptosis occurred, the content of firefly luciferase expressed in the pFluc-DEVD plasmid transfected group was about 3 times higher than pFluc-DEVG plasmid transfected group. Furthermore, Western blotting detection indicated that the Fluc level was significantly increased in pFluc-DEVD transfected group when pre-treated by apoptosis stimulants. The activation degree of Caspase-3 was closely related to the expression of Fluc, and had a significant statistical difference. These results confirmed that firefly luciferase protein expressed by pFluc-DEVD plasmid can be cleaved by the intracellular Caspase-3 enzyme, and this plasmid can accurately reflect the cell apoptosis level, which provides a useful method for quantitative detection of apoptosis.
Apoptosis ; Genes, Reporter ; Luciferases, Firefly ; Transfection

Objective To investigate the expression of T cell immunoglobulin and mucindomain-containing molecule-3 (TIM3) on PBMCs,and the plasma concentrations of soluble forms of Galetcin9 and their clinical relationship with rheumatoid arthritis (RA).Methods Peripheral blood samples were collected from 39 patients,25 osteoarthritis (OA) patients and 20 healthy subjects (HC).The expressions of TIM3 on peripheral blood mononuelear cells (PBMCs) were detected by flow cytometry.The concentrations of soluble Galetcin9 were assessed by enzyme linked immunosorbent assay (ELISA).And the relationship between their expression levels and clinical manifestations were analyzed.Levene F test was used for statistical analysis,normal distribution data were compared by t test and Pearson correlation analysis,while Mann-Whitney U test and Spearman correlation analysis were used for non-normal distribution data.Results The expression of TIM3 on CD4+ T cells was significantly higher than that of the HC [(14.7±3.2)％ vs (5.1±0.8)％,t=2.339,P=0.022 7],while there was no statistical difference between the RA group and the OA group [(14.7±3.2)％ vs (5.8±0.4)％,t=1.928,P=0.058 9].The expression of TIM3 was significantly correlated with the concentration of RF in the serum and the corresponding DAS28 score (r=0.325 8,P=0.043 0;r=0.407 5,P=0.010 0).The expression of TIM3 on CD8 + T cells in RA patients was significantly higher than that in the HC and OA [(21.1±3.4)％ vs (8.3±1.5)％,t=2.531,P=0.0142;(21.1±3.4)％ vs (10.7±1.0)％,t=2.314,P=0.024 0] which was significantly correlated with the concentration of RF in serum and the corresponding DAS28 score (r=0.451 5,P=0.003 9;r=0.524 1,P=0.000 6) as well.However,the expression of TIM3 on CD56+ NK cells was not significantly different from that of either OA or HC[(56.4±3.4)％ vs (50.6±3.8)％,t=1.047,P=0.299 8;(56.4± 3.4)％ vs (56.1±3.4)％,t=0.048,P=0.961 9],the concentration of serum RF and the corresponding DAS28 score were not significantly related.We also found that plasma Galectin 9 concentrations in RA patients were significantly higher than those of OA patients [(4.24±0.22) ng/ml vs (3.15±0.18) ng/ml,t=3.187,P=0.024] and healthy subjects [(4.24±0.22) ng/ml vs (2.55±0.14) ng/ml,t=5.567,P＜0.01],which was correlated with RF and DAS28 (r=0.479 2,P=0.002 0;r=0.353 0,P=0.027 5) while there was no correlation with CRP (r=0.176 3,P=0.283 1).Conclusion The upregulated expressions of TIM3 on peripheral lymphocytes and the high levels of plasma concentration of soluble Galectin 9 are closely correlated with the severity of the disease,suggesting that TIM3/Galectin 9 pathway may play a critical role in the pathogenesis of RA.

Objective To analyze the patients′ psychological resilience level of postpartum pelvic floor dysfunction and to explore the influencing factors, in order to provide the scientific basis for patients′psychological intervention. Methods A cross-sectional survey was conducted on 208 patients of postpartum pelvic floor dysfunction by using the Connor-Davidson Resilience scale(CD-RISC), Positive and Negative Affective scales(PANAS) and Medical Coping Modes questionnaire(MCMQ, which includes "confrontation","acceptance-resignation"and"avoidance"). Results The score of psychological resilience of 208 patients was (53.64 ± 12.29)points, differences in age, family monthly income, living locations, delivery methods and delivery times reached a statistically significant level(F=15.287,5.731,t=-2.571,-15.911,-2.922,all P<0.05). Patients′psychological resilience level had significant positive correlation with positive emotions(P<0.01)and"confrontation",and it had negative correlation with"acceptance-resignation"(P<0.01).Age,living locations, delivery methods, positive emotions and "acceptance- resignation" are the forecast factors of psychological resilience, which can explain 69.6% of total variation. Conclusions Patients of postpartum pelvic floor dysfunction have low levels of psychological resilience, which associated with positive emotions and medical coping modes. Therefore, nursing staffs should pay attentions to the patients′ psychological status, develop individualized psychological guidelines,implement psychological intervention and health education,in order to improve the level of patients′resilience and improve their living quality.

Objective To systematically evaluate the effectiveness of different nursing interventions for female overactive bladder, and to provide evidence- based evidence for nursing intervention for overactive bladder in women. Methods PubMed, Cochrane Library, Embase, ScienceDirect and CNKI,Wanfang database and CBM comprehensively by computer and included in the literature of nursing intervention for female overactive bladder patients. Results Four randomized controlled trials were included, a total of 426 participants were in, the intervention group was 214 participants, control group was 212 participants, the results of the study showed that pelvic floor muscle training,health education,psychological nursing intervention can improve patients with overactive bladder symptoms and enhance the quality of life. Conclusion Nursing intervention for women with overactive bladder is an effective way to control symptoms such as frequent micturition and urgency,improve bladder overactivity symptoms and enhance quality of life.

T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
Chickens ; Clone Cells ; Cycloheximide ; Cytoplasmic Granules ; Humans ; Protein Biosynthesis ; RNA Precursors ; RNA-Binding Proteins ; T-Lymphocytes

The cardiovascular disease has become one of the important chronic health problems for humans. At present, the research on the traditional risk factors of cardiovascular disease has been studied from the relevance to the mechanism. In this paper, the recent progress of the traditional risk factors of CVD and the impact mechanism are reviewed in order to provide a basis for the prevention, treatment and nursing of cardiovascular diseases.

Fatigue is commonly experienced in patients with advanced kidney disease and associated with poor outcomes. The purpose of this review was to discuss the prevalence, assessment methods and contributing factors of fatigue in patients with end-stage renal disease (ESRD), and also examined possible interventions to improve fatigue and concludes by defining some future research directions.