Objective:To investigate the mechanism of Fusobacterium nucleatum ( Fn) in regulating the formation of neutrophil extracellular trap (NET) to aggravate colitis. Methods:With a completely randomized design, 15 C57BL/6 mice were randomly divided into negative control group, dextran sulfate sodium (DSS) A group (DSS-induced colitis), and DSS+ Fn A group (DSS-induced colitis with Fn infection); another 15 C57BL/6 mice were randomly divided into DSS B group, DSS+ Fn B group, and GSK484 group (DSS-induced colitis with Fn infection and followed by intraperitoneal injection of peptidylarginine deiminase 4 (PAD4)inhibitor GSK484), with 5 mice in each group. The Fn-infected neutrophils (HL-60 cell) model and PAD4-inhibited cell model were established and divided into neutrophil-like control cell (induced with 1.25% dimethylsulfoxide), Fn+ neutrophil-like cell (neutrophil-like cells co-cultured with Fn), and GSK484 cell ( Fn+ neutrophil-like cell co-cultured with GSK484) with a completely randomized design. The neutrophil-like control and Fn+ neutrophil-like cells were divided into 2 batches to conduct experiments before and after PAD4 inhibition separately.The expression of zonula occludens-1 (ZO-1), neutrophil elastase (NE), reactive oxygen species, PAD4, and citrullinated histone H3(cit-H3) were analyzed by immunohistochemistry staining, Western blotting, and flow cytometry assay. Two-tailed t test was used for statistical analysis. Results:The results of immunohistochemical staining showed that compared with those of the DSS A group, the expression levels of NE, PAD4 and cit-H3 of the DSS+ Fn A group were upregulated, and the expression level of ZO-1 was downregulated; compared with those of the DSS+ Fn B group, the expression level of ZO-1 of the GSK484 group was upregulated, and the expression levels of cit-H3 and PAD4 were downregulated. The results of Western blotting demonstrated that, before the PAD4 inhibition, the expression levels of NE, PAD4 and cit-H3 of the Fn+ neutrophil-like cell were all higher than those of the neutrophil-like control cell (1.52±0.11 vs. 1.00±0.19, 1.21±0.06 vs. 1.00±0.07, 1.59±0.16 vs. 1.00±0.16), and the differences were statistically significant ( t=-4.13, -3.86, and -4.47; P=0.014, 0.014, and 0.018); after the PAD4 inhibition, the expression levels of PAD4, cit-H3, and NE of the GSK484 cell were all lower than those of the Fn+ neutrophil-like cell (0.95±0.09 vs. 1.27±0.04, 1.15±0.34 vs. 2.29±0.50, 1.22±0.14 vs. 1.68±0.12), and the differences were statistically significant ( t=5.61, 3.24, and 4.49; P=0.005, 0.032, and 0.011). The results of flow cytometry assay indicated that the positive rate of reactive oxygen species of the DSS+ Fn A group was higher than that of the DSS A group ((21.15±2.93)% vs. (11.14±1.42)%), and the positive rate of reactive oxygen species of the Fn+ neutrophil-like cell was also higher than that of the neutrophil-like control cell before the PAD4 inhibition ((51.69±6.94)% vs.(31.95±3.31)%), and the differences were statistically significant( t=5.33 and 4.45, P=0.006 and 0.011). Conclusion:Fn can promote neutrophil to release NET by upregulating reactive oxygen/PAD4/cit-H3 signaling pathway, which disrupt the intestinal barrier and aggravate colitis.

Objective:To investigate the mechanism of Fusobacterium nucleatum ( Fn) in regulating the formation of neutrophil extracellular trap (NET) to aggravate colitis. Methods:With a completely randomized design, 15 C57BL/6 mice were randomly divided into negative control group, dextran sulfate sodium (DSS) A group (DSS-induced colitis), and DSS+ Fn A group (DSS-induced colitis with Fn infection); another 15 C57BL/6 mice were randomly divided into DSS B group, DSS+ Fn B group, and GSK484 group (DSS-induced colitis with Fn infection and followed by intraperitoneal injection of peptidylarginine deiminase 4 (PAD4)inhibitor GSK484), with 5 mice in each group. The Fn-infected neutrophils (HL-60 cell) model and PAD4-inhibited cell model were established and divided into neutrophil-like control cell (induced with 1.25% dimethylsulfoxide), Fn+ neutrophil-like cell (neutrophil-like cells co-cultured with Fn), and GSK484 cell ( Fn+ neutrophil-like cell co-cultured with GSK484) with a completely randomized design. The neutrophil-like control and Fn+ neutrophil-like cells were divided into 2 batches to conduct experiments before and after PAD4 inhibition separately.The expression of zonula occludens-1 (ZO-1), neutrophil elastase (NE), reactive oxygen species, PAD4, and citrullinated histone H3(cit-H3) were analyzed by immunohistochemistry staining, Western blotting, and flow cytometry assay. Two-tailed t test was used for statistical analysis. Results:The results of immunohistochemical staining showed that compared with those of the DSS A group, the expression levels of NE, PAD4 and cit-H3 of the DSS+ Fn A group were upregulated, and the expression level of ZO-1 was downregulated; compared with those of the DSS+ Fn B group, the expression level of ZO-1 of the GSK484 group was upregulated, and the expression levels of cit-H3 and PAD4 were downregulated. The results of Western blotting demonstrated that, before the PAD4 inhibition, the expression levels of NE, PAD4 and cit-H3 of the Fn+ neutrophil-like cell were all higher than those of the neutrophil-like control cell (1.52±0.11 vs. 1.00±0.19, 1.21±0.06 vs. 1.00±0.07, 1.59±0.16 vs. 1.00±0.16), and the differences were statistically significant ( t=-4.13, -3.86, and -4.47; P=0.014, 0.014, and 0.018); after the PAD4 inhibition, the expression levels of PAD4, cit-H3, and NE of the GSK484 cell were all lower than those of the Fn+ neutrophil-like cell (0.95±0.09 vs. 1.27±0.04, 1.15±0.34 vs. 2.29±0.50, 1.22±0.14 vs. 1.68±0.12), and the differences were statistically significant ( t=5.61, 3.24, and 4.49; P=0.005, 0.032, and 0.011). The results of flow cytometry assay indicated that the positive rate of reactive oxygen species of the DSS+ Fn A group was higher than that of the DSS A group ((21.15±2.93)% vs. (11.14±1.42)%), and the positive rate of reactive oxygen species of the Fn+ neutrophil-like cell was also higher than that of the neutrophil-like control cell before the PAD4 inhibition ((51.69±6.94)% vs.(31.95±3.31)%), and the differences were statistically significant( t=5.33 and 4.45, P=0.006 and 0.011). Conclusion:Fn can promote neutrophil to release NET by upregulating reactive oxygen/PAD4/cit-H3 signaling pathway, which disrupt the intestinal barrier and aggravate colitis.

Objective To investigate the effect of 5-Aza-CdR on Notch1 pathway and neural regeneration and to explore the effects of 5-Aza-CdR on learning memory ability in mice by exploring active avoidance behavior.Methods Sixty 6～8-week-old SPF-grade ICR male mice were divided into two groups.5-Aza-CdR was administered to one group of mice via lateral ventricular injection,while the control group was injected with bovine serum albumin.Notch1 and HES1 mRNA and protein expression levels were detected by Real-time PCR and Western blot 24 hours after injection;5-bromo-2'-deoxyuridine-positive cells were observed by laser confocal microscopy,and Notch1 expression in hippocampal dentate gyrus was viewed with laser confocal microscopy.Notch1 methylation changes were detected by ethylation-specific PCR,and learning and memory behaviors of mice were assessed by passive avoidance tests and shuttle avoidance assays.Results Injection of 5-Aza-CdR increased hippocampal Notch1 pathway activity,promoted neuronal regeneration in the DG region,decreased methylation levels in the Notch1 promoter region,and enhanced the ability of mice to perform active avoidance behavior.Conclusions The effect of 5-Aza-CdR on active avoidance behavior may be related to the influence of hippocampal neural regeneration through the Notch 1 pathway.

Objective:To explore and analyze the selection of surgical methods for supratentorial intracerebral hemorrhage.Methods:A total of 260 patients with spontaneous intracerebral hemorrhage who underwent surgery in Department of Neurosurgery, Suzhou Hospital Affiliated to Nanjing Medical University from January 2017 to December 2021 were included in the study by retrospective case analysis. According to different surgical methods, they were divided into three groups: large bone flap group ( n=116), conventional bone flap group( n=89)and stereotactic group( n=55). The large bone flap group underwent standard supratentorial large bone flap craniotomy, the conventional bone flap group underwent conventional bone flap craniotomy, and the stereotactic group underwent stereotactic hematoma puncture suction + drainage. Clinical indicators such as operation time, intraoperative bleeding, pulmonary infection, length of hospital stay, and Glasgow outcome scale (GOS) at 6 months of postoperative follow-up, and the proportion of good prognosis (GOS 4-5) were calculated. Measurement data with normal distribution were expressed as mean±standard deviation( ± s), count data were expressed as cases and percentages (%). Results:In the large bone flap group, the operation time, intraoperative bleeding, hospital stay, pulmonary infection, postoperative rebleeding were(193±24) min, (625±65) mL, (46±11) d, 102 patients(87%), 9 patients(7.8%), and (124±17) min, (297±35) mL, (32±9) d, 29 patients(33%), 4 patients(4.4%)in the conventional bone flap group, and (73±11) min, (53±15) mL, (21±4) d, 10 patients(18%), 2 patients(3.6%)in stereotactic group. All patients were followed up for 6 months, and 165 patients (63.5%) had good prognosis (GOS 4-5), including 36 patients (31%) in the large bone flap group, 82 patients (93.2%) in the conventional bone flap group, and 47 patients (85.5%) in the stereotactic group.Conclusion:Standard large craniectomy has sufficient effect of decompression, and is suitable for serious life threatening hematoma; Conventional craniotomy has advantages in the treatment of secondary intracerebral hemorrhage. Stereotactic surgery has the characteristics of short operation time, less intraoperative bleeding, short hospital stay and low incidence of pulmonary infection, which is worthy of promotion in the treatment of primary intracerebral hemorrhage.

Objective To study the effect of type Ⅱ collagen (C Ⅱ) from Zaoeys dummies (Cantor)on the immune functions of rats with collagen-induced arthritis and to understand the mechanisms of RA treated with Zaoeys dhumnades (Cantor).Methods The rats with collagen-induced arthritis were randomly divided into three group:C Ⅱ from Zaocys dhumnades,bovine C Ⅱ and arthritis control group,a normal control group was set up,too.Every group had 7 rats.The C Ⅱ from Zaocys dhumnades and the bovine C Ⅱ group were fed with C Ⅱ from Zaocys dhumnades (15 mg/kg) and bovine C Ⅱ (15 mg/kg) per day respectivelyfor 15 days.The CD4/CD8 subset ratio,serum levels of anti-C Ⅱ antibody,TNF-a,IL-10,IL-1 and IL-4 in.rats were measured.Results In the arthritis group,CD4/CD8 subset ratio (P＜0.05) and serum levels of type Ⅱ collagen antibody (P＜0.01) and TNF-a (P＜0.01) were significant increased and IL-10 (P＜0.01) was significantly decreased.In the C Ⅱ from Zaocys dhumnades and bovine C Ⅱ group,CD4/CD8 subset ratio (P＜0.05) and the level of TNF-a (P＜0.01) were significantly decreased compared with the arthritis group,and had no difference compared with the normal group.The level of anti-C Ⅱ antibody was declined significantly compared with the arthritis group (P＜0.05) and had statistical difference with the normal group (P＜0.01).The level of IL-10 was significantly increased(P＜0.01),but lower than the normal group(P＜0.05).There was no statistical difference in the level of IL-1 and IL--4 in.all four groups.Conclusion C Ⅱ from Zaoeys dhum-nades (Cantor) is as effective as bovine C Ⅱ in modifying the immune functions of collagen-induced arth-ritis in rats.They can decrease the level of anti-C Ⅱ antibody,the level of TNF-a,CD4/CD8 subset ratio and increase the level of IL-10 in the peripheral blood of rats with collagen-induced arthritis.C Ⅱ from Zaocys dhumnades may be one of the important pharmacological active components that have the potential in treating rheumatoid arthritis.

The monocot mannose-binding lectin can inhibit HIV from infecting the target cells. The total RNA of Zephyranthes grandiflora was extracted and reversely transcribed into cDNA. Degenerate primers were designed based on the conserved regions of other monocot mannose-binding agglutinins by homology alignment. The 694bp full-length cDNA of Zephyranthes grandiflora agglutinin (ZGA) was cloned by RT-PCR, 3' and 5' RACE (rapid amplification of cDNA ends). The start codon and stop codon of ZGA were at 37-39bp and 529-531bp respectively. The NCBI Blast analysis result showed that ZGA gene encoded a protein precursor with signal peptide, mature protein and C-terminal cleavage sequence. The mature ZGA protein contained 106 amino acids residues and its molecular weight was 11.6KD. The percentages of identity of the deduced mature ZGA protein with those of Galanthus nivalis agglutinin, Narcissus hybrid cultivar agglutinin, Lycoris radiate agglutinin and Clivia miniata agglutinin were 71.8%, 81%, 81.8% and 84.5%, respectively. Blocks analysis revealed that ZGA had three functional domains and three mannose-binding boxes (QDNY).
Agglutinins ; genetics ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Liliaceae ; genetics ; Mannose-Binding Lectin ; genetics ; Molecular Sequence Data ; Sequence Analysis, DNA

OBJECTIVETo evaluate the efficacy and safety of leflunomide in comparison with methotrexate (MTX) on patients with rheumatoid arthritis (RA) in China.

METHODSFive hundred and sixty-six patients with active rheumatoid arthritis were randomly assigned to receive leflunomide at 20 mg once daily or MTX at 15 mg once weekly in a controlled trial. Five hundred and four patients completed the 12-week treatment and some patients continued the treatment for 24 weeks.

RESULTSBoth leflunomide and MTX could improve the symptoms, signs, and joint function, but there were no changes in X-ray observations of patients with rheumatoid arthritis. In the leflunomide group, the overall rates of effectiveness at 12 weeks and 24 weeks were 86.94% and 92.31% respectively; the rates of remarkable improvement were 64.95% and 79.81% respectively. In the MTX group, the overall rates of effectiveness at 12 weeks and 24 weeks were 84.04% and 83.15% respectively; the rates of remarkable improvement were 56.81% and 75.28% respectively. According to intent-to-treat analysis, the ACR 20% response rates at 12 weeks and 24 weeks in the leflunomide group were 62.54% and 67.18% respectively, compared with 60.08% and 61.32% respectively in MTX group. No statistical differences were shown in the efficacy between the two groups (P > 0.05). The adverse events in the leflunomide group were gastrointestinal symptoms, skin rash, alopecia, nervous system symptoms, decreased leukocyte count, and elevation of alanine aminotransferase (ALT). Most of these side effects were mild and transient. The incidence of adverse events in the leflunomide group was 16.84%, significantly lower than that in MTX group (28.17%, P = 0.002).

CONCLUSIONSLeflunomide is effective in the treatment of RA with less adverse events than MTX. Its efficacy is similar to MTX, but the incidence of adverse events and the rate of withdrawal due to adverse events were lower in the leflunomide group than in MTX group.

Antirheumatic Agents ; adverse effects ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Female ; Growth Inhibitors ; adverse effects ; therapeutic use ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Isoxazoles ; adverse effects ; therapeutic use ; Male ; Methotrexate ; adverse effects ; therapeutic use ; Middle Aged