Objective To investigate changes in gut microbiota during diabetic nephropathy(DN)pro-gression using 16S rDNA sequencing technology.Methods A total of 90 male SD rats were randomly divided into a normal control group(n=10,no modeling,regular feeding)and a model group(diabetes model).The diabetes model was established by a single intraperitoneal injection of streptozotocin(STZ)at 60 mg/kg,with regular feeding.According to the feeding time after modeling,the rats were divided into 2-week,4-week,8-week,and 12-week model groups(fed for 2,4,8,and 12 weeks after model establishment),with 20 rats in each group.Blood urea nitrogen(BUN)was measured using the urease method,serum creatinine(Scr)was deter-mined by the picric acid method,and ELISA was used to detect urinary kidney injury molecule-1(KIM-1)and neutrophil gelatinase-associated lipocalin(NGAL)levels.HE,PAS,and Masson staining were used to observe renal tissue pathological changes.Gut microbiota was collected from the rats,and 16S rDNA gene sequencing was used to analyze the gut microbiota to understand changes in the gut microbiota.Results Compared with the normal control group,the levels of KIM-1 and NGAL in urine of rats in all model groups were significantly increased(P<0.05).Pathological staining results showed that,compared with the normal control group,rats in all model groups exhibited diffuse thickening of the glomerular basement membrane and pathological chan-ges such as local necrosis and vacuolar degeneration in renal tubular epithelial cells.16S rDNA sequencing re-sults indicated that the abundance and structure of intestinal microbiota in rats of all model groups changed.Compared with the normal control group,in the 8-week and 12-week model groups,the relative abundance of Bacteroides and Akkermansia decreased,while the relative abundance of Roseburia,Alloprevotella,Prevotel-laceae-Ga6A1,and Ruminococcaceae UCG-005 increased.Compared with the normal control group,in the 12-week model group,the abundance of Akkermansia decreased and that of Prevotellaceae-NK3B31 increased.Conclusion The abundance and structure of gut microbial community in DN rats under conventional feeding at different time points change significantly,further confirming the"gut-kidney axis"theory.

Objective:To explore the effect of comfort nursing intervention in hospice care for advanced cancer patients at home, and to provide reference for hospice care for advanced cancer patients at home.Methods:A randomized controlled trial was conducted. A total of 105 patients with advanced cancer who were treated in the Cancer Hospital Affiliated to Harbin Medical University from January to February 2023 were selected as the research objects. According to the random number table, they were divided into control group of 53 cases and intervention group of 52 cases. The control group received routine nursing methods, while the intervention group received comfort nursing interventions on this basis. The intervention lasted for 4 weeks. The changes in palliative care outcomes, quality of life and death anxiety were compared between the two groups before and after intervention.Results:Totally 105 cases were included, 53 cases in the control group and 52 cases in the intervention group. In the control group, there were 25 males, 28 females, aged (58.96 ± 10.71) years old; in the intervention group, there were 22 males, 30 females, aged (59.82 ± 10.53) years old. Before intervention, there was no statistically significant difference in palliative care outcomes, quality of life and cancer death anxiety scores between the two groups of patients (all P>0.05). After intervention, the total score of palliative care outcomes in the intervention group was (13.34 ± 5.88) points, significantly lower than (16.15 ± 5.72) points in the control group, with a statistically significant difference ( t = 2.48, P<0.05). The overall health status score of quality of life was (68.55 ± 9.34) points in the intervention group, higher than (63.01 ± 9.28) points in the control group ( t = 3.05, P<0.05). The total score of cancer death anxiety was (8.85 ± 2.72) points, significantly lower than (10.59 ± 3.14) points of the control group, with a statistically significant difference ( t = 3.04, P<0.05). Conclusions:The application of comfort nursing mode in home based hospice care can improve the quality of hospice care for advanced cancer patients, reduce their level of death anxiety, and is of great significance for improving the quality of life of advanced cancer patients.

Objective To explore the mechanism of curcumin in enhancing gemcitabine sensitivity by regulating pancreatic cancer cells.Methods In order to verify the effect of curcumin on p53 and NF-κB p65 nuclear translocation in pancreatic cancer cell PANC1,the pancreatic cancer PANC1 cells were treated with 0,10,20,30 μmol/L curcumin to obtain the blank group,10 μmol/L curcumin group,20 μmol/L curcumin group and 30 μmol/L curcumin group.The intracellular distribution of NF-κB p65 was determined by immunofluo-rescence staining under fluorescence microscope.The expression level of p53 protein in cells was determined by Western blot.The expression level of p53 mRNA was determined by reverse transcription-real-time fluo-rescence quantitative PCR.The combined use of miRstar and JASPAR softwares predicted to seek microRNA (miRNA) potentially regulated by NF-κB p65 nuclear translocation.The double luciferase reporting assay was used to respectively verify the relationship between miRNA and p53.In order to verify whether curcumin in-fect the sensitivity of PANC1 cell through NF-κB p65/miR-266-5p/p53 axis,the gemcitabine group was ob-tained by 10 μmol/L gemcitabine treating cells,and the curcumin combined gemcitabine group was obtained by 20 μmol/L curcumin and 10 μmol/L gemcitabine treating cells.After the cells were treated with curcumin (20 μmol/L) and gemcitabine (10 μmol/L),oligonucleotides mimic NC and miR-26b-5p mimic were trans-fected into cells,and the mimic NC group and miR-26b-5p group were obtained.pcDNA3.1 no-load plasmid and pcDNA3.1-p53 overexpression plasmid were transfected into cells,respectively,and the pcDNA3.1 group and p53 group were obtained.The cell activity was detected by MTT assay.The cellular apoptosis level was determined by Annexin V/PI double staining.Results Compared with the blank group,the intracellular p53 mRNA and protein expression levels in the various curcumin treated groups (10,20,30 μmol/L curcumin groups) all were increased.Compared with the blank group,the expression level of NF-κB p65 in the nucleus of 20μmol/L curcumin group was decreased.The miRstar and JASPAR prediction software found 8 miRNA that may be regulated by NF-κB p65 nuclear translocation,in which 3 miRNA had potential to target p53 genes,especially miR-26b-5p effect was most significant.The double luciferase report experiment confirmed that miR-26b-5p did interact with p53.Compared with the mimic NC group,the expression levels of p53 mR-NA and protein in the mimic miR-26b-5p group were decreased.Compared with the gemcitabine group,the cell viability in the curcumin combined gemcitabine group was decreased and the apoptosis rate was increased.Compared with the mimic NC group,the cell activity in the mimic miR-26b-5p group was increased and the apoptosis level was decreased.Moreover compared with the pcDNA3.1 group,the cell activity in the pcDNA3.1-p53 group was decreased and the apoptosis level was increased.Conclusion Curcumin reduces the expression of miR-26b-5p gene by inhibiting the nuclear translocation of NF-κB p65 protein,and then increase the p53 gene and protein expression,and enhance the sensitivity of pancreatic cancer cells to gemcitabine by the NF-κB p65/miR-26b-5p/p53 axis.

Objective:To investigate the effects of induced pluripotent stem cell-derived exosome (iPSC-Exo) on releasing inflammatory factors from microglia induced by lipopolysaccharide (LPS).Methods:iPSC derived from the tubular epithelial cells of sepsis encephalopathy patients were resuscitated and cultured. The iPSC-Exo was isolated by low-temperature ultracentrifugation and analyzed by transmission electron microscopy, Western blot and high sensitivity flow cytometry (HSFCM). Based on the concentration of iPSC-Exo, human microglia line HMO6 cells activated by LPS (100 ng/mL) were divided into four groups randomly: LPS+ phosphate buffer solution (PBS) group, LPS+iPSC-Exo 10 5 group, LPS+iPSC-Exo 10 6 group and LPS+iPSC-Exo 10 7 group. The control group was added equal PBS but not LPS. After culture for 24 h, the concentrations of malondialdehyde in cells were detected. Quantitative RT-PCR was used to measure the mRNA expression levels of macrophage inflammatory protein 2 (MIP2), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 in the cells and enzyme-linked immunosorbent assay (ELISA) was used to assess the concentration of these cytokines in the supernatant. Under the same concentration of iPSC-Exo, one-way ANOVA and SNK- q test were used for comparison between groups. Results:The extracts showed spherical membrane structure by transmission electron microscopy. HSFCM showed the mean diameter of the extracts was (74.66±15.60) nm and the concentration around 2.98×10 10/mL. Western blot analysis showed high expression of exosome markers CD63, Alix and TSG101, but not GM130. Intracellular MDA concentration and mRNA expression levels and protein concentration of MIP2, TNF-α, IL-1β and IL-6 in the LPS+PBS group were significantly higher than those in the control group (all P<0.01). With the increase of iPSC-Exo concentration, the intracellular MDA concentration decreased gradually ( P<0.01), the mRNA expression levels of inflammatory factors showed a gradual downward trend (all P<0.05). Each inflammatory cytokine in the supernatant declined in a manner that was almost consistent with mRNA. Concentrations of MDA remained constant in the control group. Conclusions:iPSC-Exo derived from the tubular epithelial cells of sepsis encephalopathy patients alleviate oxidative stress and inflammation effect of microglia induced by LPS, and the modulatory effect is dose-dependent.

OBJECTIVE: To analyze the composition and metabolites of gut microbiota in septic rats by fecal 16s rRNA sequencing and untargeted metabolomics, and to preliminarily explore the effect and potential mechanism of gut microbiota and its metabolites on inflammatory response and multiple organ damage in sepsis. METHODS: Ten males healthy male Wistar rats were randomly divided into a sham operated group (Sham group) and sepsis model group (CLP group) using a random number table method, with 5 rats in each group. A rat sepsis model was established by cecal ligation and perforation (CLP) method. The animals were sacrificed 24 hours after modeling, the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung and kidney tissues, and the pathological scores were evaluated. Fecal samples were collected, and 16s rRNA high-throughput sequencing and non-targeted metabolomics were used to screen microbiota, metabolites and potential signal pathways that may play an important role in disease outcomes. Spearman correlation analysis was conducted to jointly analyze the gut microbiota and non-targeted metabolism. RESULTS: Compared with the Sham group, the degree of pathological damage to lung and kidney tissues in the CLP group was significantly increased (lung tissue score: 3.60±0.80 vs. 0.00±0.00, kidney tissue score: 2.40±0.80 vs. 0.00±0.00, both P < 0.01), the level of IL-6 and TNF-α in peripheral blood significantly increased [TNF-α (ng/L): 248.12±55.98 vs. 143.28±36.57, IL-6 (ng/L): 260.26±39.47 vs. 116.01±26.43, both P < 0.05], the species diversity of intestinal flora of rats in the CLP group was significantly reduced, the relative abundance of Morganella, Bacteroides and Escherichia-Shigella were significantly increased, and the relative abundance of Lachnospiraceae NK4A136, Ruminococcus, Romboutsia and Roseburia were significantly reduced. In addition, the biosynthesis and bile secretion of phenylalanine, tyrosine, and tryptophan in the gut microbiota of the CLP group were significantly increased, while the biosynthesis of secondary bile acids was significantly reduced. There was a significant correlation between differential metabolites and differential microbiota. CONCLUSIONS Sepsis can cause significant changes in the characteristics of gut microbiota and fecal metabolites in rats, which provides a basis for translational research to seek new targets for the treatment of sepsis.
Rats ; Male ; Animals ; Tumor Necrosis Factor-alpha ; RNA, Ribosomal, 16S ; Gastrointestinal Microbiome ; Interleukin-6 ; Rats, Wistar ; Sepsis

OBJECTIVE：To investigate the effect of increasing efficacy and decreasing toxicity of Limax extract （LE）on cyclophosphamide（CTX）in the treatment of hepatocellular carcinoma. METHODS ：The mice were randomly divided into normal group，model group ，CTX group （0.02 g/kg），LE low-dose ，medium-dose and high-dose groups （LEL，LEM，LEH group ，0.6，1.2，2.4 g/kg），CTX+LE low-dose ，medium-dose and high-dose combination groups （CTX+LEL，CTX+LEM，CTX+ LEH group ，the same dose as single drug group ），with 10 huangrenbin518@163.com mice in each group. Except for normal group ，other groups were inoculated with hepatoma cells H 22 in the left ar mpit to establish tumor bearing models. After 24 h of inoculation ，normal group and model group were intragastrically given normal saline ， and administration groups were intragastrically given corresponding drugs ，once a day ，for 10 days. On the second day after the last administration ，the general conditions of mice in each group were observed ；the body mass ，thymus index （LI），spleen index （SI）were measured ；the tumor inhibition rate was detected. The effect （q）of combination therapy was evaluated by King ’s formula . The counts of WBC ，RBC and PLT ，serum contents of ALT ，ALT，Scr and BUN were detected in model group ，CTX group and combination groups ，and the contents of MDA，SOD and GSH ，the levels of VEGF ，TNF-α and IL-6 in the tumor tissue were detected by colorimetry and ELISA in above groups. The protein expression of oncogenes （p53，Bcl-2 and Bax ）were detected by immunohistochemical method in model group，CTX group and CTX+LEM group. RESULTS ：The mice in the model group were in poor spirit and had symptoms of excessive drinking and eating ；although the body weight ，TI and SI were not significantly abnormal compared with normal group （P＞0.05），WBC count and AST content were significantly increased ，ALT and BUN contents were significantly decreased （P＜ 0.05 or P＜0.01）. Compared with model group ，above symptoms of mice were all improved in administration groups. The tumor weight of administration groups ，TI and SI of CTX group and TI of combination groups were decreased significantly ，but tumor weight of LEL group and LEH group ，TI and SI of LE single groups and combination groups were significantly higher than CTX group；tumor weight of combination groups were significantly lower than CTX group （P＜0.05 or P＜0.01）. The tumor inhibition rates of administration groups were 29.58％-72.08％. The q values of CTX+LEL group ，CTX+LEM group and CTX+LEH group were 1.03，0.97 and 0.86，respectively. Compared with model group ，WBC count ，AST and BUN contents of CTX group ，MDA contents of combination groups ，VEGF，TNF-α and IL-6 levels of administration groups ，the protein expression of Bcl- 2 in CTX group and CTX+LEM group were decreased significantly ；the activities of SOD and GSH of administration groups ，the protein expression of p 53 in CTX+LEM group and Bax in CTX group ，CTX+LEM group were increased significantly （P＜0.05 or P＜ 0.01）；WBC counts and AST contents of administration groups ，ALT content of CTX+LEM group ，SOD activity of CTX+LEH group and GSH activity of CTX+LEM group were all significantly higher than those of CTX group ；MDA content of CTX+LEH group，VEGF and TNF-α levels of CTX+LEM group and CTX+LEH group，IL-6 levels of administration groups were all significantly lower than CTX group （P＜0.05 or P＜0.01）. CONCLUSIONS ：LE combined with CTX can increase the anti-tumor effect，and LE can reduce the toxicity of CTX induced immunosuppression and bone marrow suppression in mice ，with effect of increasing efficacy and decreasing toxicity. The effect may be related to antioxidant stress ，inhibition of angiogenesis and secretion of inflammatory factors ，and regulation of apoptosis protein expression.

Objective: To investigate the relationship between ultrasound derived ratio of femoral vein to femoral artery diameter and hemodynamics in patients with heart failure. Methods: This was a case-control study. A total of 61 patients with heart failure and 49 patients with non-heart failure hospitalized in the Department of Critical Care Medicine from September 2017 to September 2018 were included in this study. Doppler ultrasound was used to measure the femoral artery and vein diameter. After deep inhalation, the femoral vein diameter was measured again, and the ratio of femoral vein and artery diameter was calculated. The central venous pressure (CVP) and mean pulmonary wedge pressure (mPAWP) were also measured. Pearson correlation analysis was used to explore the correlation between the ratio of femoral vein diameter to femoral artery diameter and CVP and mPAWP, and linear regression equation was established. Results: The overall CVP and mPAWP levels were significantly higher, and the femoral vein diameter after deep inhalation was bigger in heart failure patients than in non-heart failure patients(all P<0.001). The femoral vein diameter/femoral artery diameter ratio was positively correlated with CVP (r=0.76, P<0.001), and positively correlated with mPAWP (r=0.40, P<0.001) in heart failure group. The linear regression equation established by the femoral vein/femoral artery diameter ratio and CVP in the heart failure group showed that the inner diameter of the femoral vein/the inner diameter of the femoral artery ratio≥1.3 corresponded CVP≥15.518 cmH₂O(1 cmH₂O=0.098 kPa) in heart failure patients. Conclusions In patients with heart failure, the inner diameter of the femoral vein/femoral artery ratio is positively correlated with CVP and mPAWP. The ratio of inner diameter of the femoral vein/femoral artery can be used to assess the volumetric load of patients with heart failure and to guide the clinical treatment of heart failure patients.

This article illuminates the status quo, shortcomings and prospects of follow-up visit at home and abroad by retrieving and summarizing the related studies on follow-up visit for enterostomy at home and abroad. It reviews the nurse-led follow-up visit from the perspective of continued nursing care, in order to provide a reference for China to further standardize the follow-up content for patients with enterostomy.

ObjectiveTo analyze the literature characteristics on essential hypertension treated by external therapy of traditional Chinese medicine, summarize the current research situation and trend in the field, and to provide a reference for relative researches.MethodsThe papers relevant to treatingessential hypertension with external therapy of TCM included inSinoMedwere statistically analyzed from the aspects of publishing year, journals distribution,author’s unit and districtdistribution,research funds and literature content with bibliometrics method.ResultsThe total number of the literature for analyzing was 226, the number of papers increased gradually.Authors of the papers were mainly from TCM universities or colleges.Papersdistributedmainly in the more economically developed regions,which issued the largest amountinGuangdongprovince.Papers supported by research funds accounted for 16.81% in all the literature. Most literaturewasclinical research and the most commonly used for external therapywasacupuncture.ConclusionThe research and clinical work of essential hypertension treatment with external therapy of traditional Chinese medicinewerepaid more close attention in recent years, but therewerestill some problems that need to be solvedto form a viable, effective treatment system.

Objective To analyze the correlation between critical thinking ability and self- efficacy for nursing students in university of traditional Chinese medicine, so as to provide references and evidences for nursing quality teaching. Methods A total of 291 nursing students in university of traditional Chinese medicine were investigated with Critical Thinking Disposition Inventory- Chinese Version (CTDI- CV) and General Self- Efficacy Scale (GSES). Results The total scores of critical thinking ability and self- efficacy were (288.24±32.95) and (25.44±5.74). There were consistence between critical thinking ability and self- efficacy (r=0.348, P<0.01). Regression analysis revealed that self- efficacy and interpersonal relations were predictors of critical thinking ability. Conclusions There were consistence between critical thinking ability and self- efficacy. Self- efficacy and interpersonal relations were predictors of critical thinking ability. It is suggested to promote critical thinking ability by enhancing self- efficacy and interpersonal relations, which contributes to improvement of comprehensive quality.