Objective:To investigate the protective effect and its mechanism of proscillaridin A (PSD-A) on podocyte injury in lupus nephritis (LN).Methods:Molecular docking and surface plasmon resonance techniques were used to analyze the binding status of PSD-A to signal transducer and activator of transcription 1 (STAT1). The immortalized human podocyte injury model in the lupus group was induced by the serum of systemic lupus erythematosus patients, and the control and PSD-A intervention (2 nmol/L, 4 nmol/L) groups were also set up. Six female 12-week-old C57BL/6 mice were designated as the control group, and 12 female 12-week-old MRL/lpr lupus mice were randomly divided into lupus group and PSD-A intervention group by random number table method. The PSD-A intervention group was intraperitoneally injected with 5 mg/kg PSD-A, once per week for 6 consecutive weeks. While the control group and the lupus group were intraperitoneally injected with the same volume of the solvent without PSD-A. Western blotting and real-time quantitative PCR were employed to detect the relative protein and mRNA expression levels of podocin, STAT1, and interferon-induced protein with tetratricopeptide repeat 1 (IFIT1) in podocytes of each group. Enzyme-linked immunosorbent assay was used to detect the levels of serum anti-double strand DNA antibody and interferon-α in mice. Coomassie brilliant blue was used to detect the urinary protein level. HE, PAS, Masson and PASM staining and transmission electron microscopy were used to observe the pathological changes of renal tissues. Immunohistochemistry was used to examine the protein expression of podocin, STAT1 and IFIT1 in renal tissues.Results:Molecular docking and surface plasmon resonance techniques proved that PSD-A could bind to STAT1 protein and they exhibited a robust binding affinity. The podocyte experiments showed that, compared with the lupus group, the relative expression levels of podocin protein and mRNA in the PSD-A intervention group were upregulated, while the relative expression levels of STAT1 and IFIT1 protein and mRNA were downregulated (all P<0.05). The animal experiments showed that, compared with the lupus group, the serum levels of anti-double strand DNA antibody, interferon-α, and urinary protein in PSD-A intervention group were decreased, the pathological damage of renal tissues was alleviated, and the injury of renal podocytes was reduced. Immunohistochemical staining showed that the relative protein expression levels of STAT1 and IFIT1 of renal tissues in the PSD-A intervention group were lower than those in the lupus group (all P<0.05). Conclusion:PSD-A can play a protective role in podocyte injury in LN, and its mechanism may be related to the inhibition of the STAT1 signaling pathway.

Objective To explore the internal correlation mechanism among symptoms and influencing factors in first-ever subacute stroke patients.Methods A cross-sectional survey method was employed to conveniently select stroke patients hospitalized in a tertiary hospital in Guangdong Province from October 2022 to June 2023.Data were collected using a general information questionnaire,Symptom Experience Scale for Stroke Survivors,Fear of Progression Questionnaire-Short Form,Acceptance of Illness Scale,Herth Hope Index,and the Memorial University of Newfoundland Scale of Happiness.Results A total of 316 stroke patients were included.The incidence of symptom burden ranged from 17.41％to 87.97％,with the core,bridge,and sentinel symptoms as decreased attention and restricted limb movements,restricted limb movements and decreased self-care ability,and limb weakness and delayed response,respectively.Age,stroke type,disease stage,self-care ability,fear of disease progression,hope level,and happiness level were predictive factors for symptom burden in patients.Mixed network analysis showed direct correlation between foot inversion and stroke type,fear of disease progression was directly associated with limb weakness and limb pain.Low mood was directly correlated with hope level and happiness level.Conclusion Symptoms in patients with first-episode subacute stroke are interrelated.Nurses should pay attention to the identification of core,bridge,and sentinel symptoms,understand the associations between symptoms and influencing factors,and provide comprehensive and personalized clinical symptom management strategies for patients.

Objective To explore the internal correlation mechanism among symptoms and influencing factors in first-ever subacute stroke patients.Methods A cross-sectional survey method was employed to conveniently select stroke patients hospitalized in a tertiary hospital in Guangdong Province from October 2022 to June 2023.Data were collected using a general information questionnaire,Symptom Experience Scale for Stroke Survivors,Fear of Progression Questionnaire-Short Form,Acceptance of Illness Scale,Herth Hope Index,and the Memorial University of Newfoundland Scale of Happiness.Results A total of 316 stroke patients were included.The incidence of symptom burden ranged from 17.41％to 87.97％,with the core,bridge,and sentinel symptoms as decreased attention and restricted limb movements,restricted limb movements and decreased self-care ability,and limb weakness and delayed response,respectively.Age,stroke type,disease stage,self-care ability,fear of disease progression,hope level,and happiness level were predictive factors for symptom burden in patients.Mixed network analysis showed direct correlation between foot inversion and stroke type,fear of disease progression was directly associated with limb weakness and limb pain.Low mood was directly correlated with hope level and happiness level.Conclusion Symptoms in patients with first-episode subacute stroke are interrelated.Nurses should pay attention to the identification of core,bridge,and sentinel symptoms,understand the associations between symptoms and influencing factors,and provide comprehensive and personalized clinical symptom management strategies for patients.

Objective:To investigate the protective effect and its mechanism of proscillaridin A (PSD-A) on podocyte injury in lupus nephritis (LN).Methods:Molecular docking and surface plasmon resonance techniques were used to analyze the binding status of PSD-A to signal transducer and activator of transcription 1 (STAT1). The immortalized human podocyte injury model in the lupus group was induced by the serum of systemic lupus erythematosus patients, and the control and PSD-A intervention (2 nmol/L, 4 nmol/L) groups were also set up. Six female 12-week-old C57BL/6 mice were designated as the control group, and 12 female 12-week-old MRL/lpr lupus mice were randomly divided into lupus group and PSD-A intervention group by random number table method. The PSD-A intervention group was intraperitoneally injected with 5 mg/kg PSD-A, once per week for 6 consecutive weeks. While the control group and the lupus group were intraperitoneally injected with the same volume of the solvent without PSD-A. Western blotting and real-time quantitative PCR were employed to detect the relative protein and mRNA expression levels of podocin, STAT1, and interferon-induced protein with tetratricopeptide repeat 1 (IFIT1) in podocytes of each group. Enzyme-linked immunosorbent assay was used to detect the levels of serum anti-double strand DNA antibody and interferon-α in mice. Coomassie brilliant blue was used to detect the urinary protein level. HE, PAS, Masson and PASM staining and transmission electron microscopy were used to observe the pathological changes of renal tissues. Immunohistochemistry was used to examine the protein expression of podocin, STAT1 and IFIT1 in renal tissues.Results:Molecular docking and surface plasmon resonance techniques proved that PSD-A could bind to STAT1 protein and they exhibited a robust binding affinity. The podocyte experiments showed that, compared with the lupus group, the relative expression levels of podocin protein and mRNA in the PSD-A intervention group were upregulated, while the relative expression levels of STAT1 and IFIT1 protein and mRNA were downregulated (all P<0.05). The animal experiments showed that, compared with the lupus group, the serum levels of anti-double strand DNA antibody, interferon-α, and urinary protein in PSD-A intervention group were decreased, the pathological damage of renal tissues was alleviated, and the injury of renal podocytes was reduced. Immunohistochemical staining showed that the relative protein expression levels of STAT1 and IFIT1 of renal tissues in the PSD-A intervention group were lower than those in the lupus group (all P<0.05). Conclusion:PSD-A can play a protective role in podocyte injury in LN, and its mechanism may be related to the inhibition of the STAT1 signaling pathway.

ObjectiveTo obtain HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect by transfecting HSC-T6 cells with CRISPR/Cas9 lentiviral vector, and to provide a good method for further functional research and new strategies for the clinical treatment of liver fibrosis. MethodsThe COX-2 gene-specific sgRNAs (COX-2-sgRNA-1, COX-2-sgRNA-2, COX-2-sgRNA-3) were designed, synthesized, and connected to the GV371 vector, and the recombinant plasmid and the packaging plasmid were transfected into 293T cells to form lentivirus particles; the fluorescence method was used to measure virus titer. The most appropriate amount of the virus was calculated based on MOI. Lenti-Cas9-puro was transfected into HSC-T6 cells, and HSC-T6-Cas9 cells were screened out by puromycin; Lenti-COX-2-sgRNA-EGFP was transfected into HSC-T6-Cas9 cells to obtain HSC-T6-COX-2-/- cells. Cruiser enzyme digestion and Western blot were used to verify gene knockout at the gene and protein levels. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsSequencing verified that the COX-2-sgRNA expression vector was constructed successfully. Recombinant expression plasmids and packaging plasmids were transfected into 293T cells to form lentivirus particles, and the fluorescence method showed a virus titer of ＞1×108. HSC-T6 cells with stable expression of Cas9 protein and HSC-T6-COX-2-/- cells with COX-2 gene defect were successfully constructed. The HSC-T6-Cas9 group had significantly higher relative mRNA expression of LV-Cas9-Puro than the CON group (541.93±105.76 vs 1.00±0.02, t=12.995, P＜0.01). Cruiser enzyme digestion and Western blot showed that the CRISPR/Cas9 lentivirus expression vectors played a role in the target, among which COX-2-sgRNA-2 knockout had the most significant effect, and this group had a significant reduction in the protein expression level of COX-2 compared with the CON group and the NC group (both P＜0.05), suggesting that COX-2-sgRNA was active. ConclusionA CRISPR/Cas9 lentivirus vector is successfully constructed for COX-2 target gene, and HSC-T6-COX-2-/- cells with stable COX-2 gene knockout are obtained.

ObjectiveTo investigate the effect and significance of cyclooxygenase-2 (COX-2) inhibitors on the expression of the Acsl gene family in the ileum of rats with nonalcoholic fatty liver disease (NAFLD). MethodsA total of 45 Sprague-Dawley rats were randomly divided into normal control group (15 rats given normal diet), NAFLD model group (15 rats given high-fat diet), and nimesulide group (15 rats given high-fat diet and nimesulide). All rats were sacrificed after 12 weeks of feeding, and then blood samples were collected from the inferior vena cava to measure total cholesterol (TC) and triglyceride (TG). HE staining and oil red O staining were performed for the liver to evaluate the degree of hepatic steatosis in each group, and quantitative real-time PCR was used to measure the mRNA expression of the Acsl family genes in the ileum. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in serum TC and TG and marked hepatic steatosis (all P＜0.05); compared with the NAFLD model group, the nimesulide group had significant reductions in serum TC and TG and degree of hepatic steatosis (all P＜0.05). Compared with the normal control group, the NAFLD model group had a significant increase in the expression of COX-2 in the ileum (P＜0.05), and compared with the NAFLD model group, the nimesulide group had a significant reduction in the expression of COX-2 in the ileum (P＜005). Compared with the normal control group, the NAFLD model group had significant increases in the mRNA expression of Acsl3 and Acsl5 in the ileum (both P＜0.05), and compared with the NAFLD model group, the nimesulide group had significant reductions in the mRNA expression of Acsl3 and Acsl5 (both P＜0.05). ConclusionThe COX-2 inhibitor nimesulide can regulate the expression of the Acsl gene family in the ileum of rats with NAFLD, suggesting that COX-2 inhibitors may inhibit the progression of NAFLD through the Acsl gene.

Background and purpose: miR-21 is ovexpressed in various types of human cancers. This study was designed to investigate the effect of miR-21 knockdown on cell proliferation, migration and invasion of nasopharyngeal carcinoma (NPC) cell line CNE2. Methods: CNE2 was transfected with miR-21 inhibitor by LipofectamineTM2000. Meanwhile CNE2 was transfected with NC inhibitor as negative control. qRT-PCR was used to detect the miR-21 expression in these cells. The effects of miR-21 downregulation on cell proliferation, migration and invasion were evaluated by MTS, wound-healing Transwell and invasion assays. Results: miR-21 expression was remarkably downregulated in miR-21 inhibitor-transfected cells in concentration-dependent manner, indicating transfection with miR-21 inhibitor can effectively reduce expression level of miR-21 in CNE2 cells. Transfection of miR-21 inhibitor into CNE2 cells led to a signiifcant decrease in cell proliferation rate compared with control cells (P<0.05). miR-21 downregulation results in reduction of cell migration(P<0.05). Moreover, the cell invasion by Transwell invasion assay was reduced in miR-21-downregulated cells relative to control cells. Conclusion:miR-21 can promote cell proliferation, migration and invasion of NPC cells. And it maybe plays an important role in tumorigenesis and development of NPC.

Objective To explore the mechanism and therapeutic effects of wet dressing with 50％ magnesium sulfate ( MgSO4 ) as a part of comprehensive treatment for wound resulted from pit viper bites in children.Methods A total of 61 children with pit-viper-bitten wound enrolled from May 2009 through May 2011 were divided into two groups,namely the comprehensive treatment group (n =31 cases) and the conventional treatment group (n =30 eases). In the comprehensive treatment group,50％ magnesium sulfate wet dressing applied to the swelling parts of body in the early stage in addition to the conventional treatment and topically bandaged once a day keeping wet with spraying saline on the top of dressing.Before the treatment and on the 1st day,the 3rd day and the 5th day of treatment,the values of myocardial enzyme and the circumference of swelling parts were recorded,and the differences were analyzed by t- test.The differences in rate of wound healing in one week between two groups were analyzed by chi-square test.Results Before the treatment,there was no statistical difference in clinical features between the two groups with different modes of treatment for the viper-bitten children (P ＞ 0.05 ). During comprehensive treatment,the values of myocardial enzyme and the circumference of swelling parts decreased more markedly than those in conventional treatment group ( P ＜ 0.05 ).The rates of wound healing in one week of two groups were 80.5％ and 56.6％ respectively.Conclusions The comprehensive treatment with adjuvant of 50％ MgSO4 for the viper-bitten children can effectively reduce the tissue damage caused by venom,and the swelling as well,restoring function of body and shortening the time for wound healing and preventing complications.