Mechanism of intestinal injury induced by WNT2B high-expressed fibroblasts in Crohn's disease / 中华儿科杂志

Objective: To explore the mechanism of intestinal tissue damage induced by macrophages activated by WNT2B high-expressed fibroblasts. Methods: This study involved biological information analysis, pathological tissue research and cell experimental research. The biological information of the colon tissue from the children with inflammatory bowel disease in previous study was analyzed again with single-cell sequencing. The pathological tissues were collected by colonoscopy from 10 children with Crohn's disease treated in the Department of Gastroenterology of Guangzhou Women and Children's Medical Center from July 2022 to September 2022. According to the findings of colonoscopy, tissues with obvious inflammation or ulceration were classified as the inflammatory group, while tissues with slight inflammation and no ulceration were classified as the non-inflammatory group. HE staining was performed to observe the pathological changes of the colon tissues. Macrophage infiltration and CXCL12 expression were detected by immunofluorescence. In terms of cell experiments, fibroblasts transfected with WNT2B plasmid or empty plasmid were co-cultured with salinomycin treated or non-treated macrophages, respectively; the expression of proteins through Wnt classical pathway were detected by western blotting. Macrophages treated with SKL2001 were used as the experimental group, and those with phosphate buffer as the control group. The expression and secretion of CXCL12 in macrophages were detected by quantitative Real-time PCR and enzyme-linked immunosorbent assay (ELISA). T-test or rank sum test were used for the comparison between groups. Results: Single-cell sequencing analysis suggested that macrophages were the main cells in inflammatory bowel disease colon tissue, and there was interaction between WNT2B high-expressed fibroblasts and macrophages. HE staining of the 10 patients ((9.3±3.8) years old, 7 males and 3 females) showed that the pathological score of colon tissue in the inflammatory group was higher than that in the non-inflammatory group (4 (3, 4) vs. 2 (1, 2) points, Z=3.05, P=0.002). Tissue immunofluorescence indicated that the number of infiltrating macrophages in the inflammatory group was significantly higher than that in the non-inflammatory group under high power field of view (72.8±10.4 vs.8.4±3.5, t=25.10, P<0.001), as well as the number of cells expressing CXCL12 (14.0±3.5 vs. 4.7±1.9, t=14.68, P<0.001). In cell experiments, western blotting suggested an elevated level of glycogen synthase kinase-3β phosphorylation in macrophages co-cultured with fibroblast transfected with WNT2B plasmid, and salinmycin could reverse this change. Real-time PCR suggested that the transcription level of CXCL12 in the experimental group was higher than that in the control group (6.42±0.04 vs. 1.00±0.03, t=183.00, P<0.001), as well as the expression and secretion of CXCL12 by ELISA ((465±34) vs. (77±9) ng/L, t=13.21, P=0.006). Conclusion: WNT2B high-expressed fibroblasts can secrete WNT2B protein and activate the Wnt classical signaling pathway thus enhancing the expression and secretion of CXCL12 in macrophages, inducing the development of intestinal inflammation of Crohn's disease.

Fibroblasts overpressing WNT2b cause impairment of intestinal mucosal barrier / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which fibroblasts with high WNT2b expression causes intestinal mucosa barrier disruption and promote the progression of inflammatory bowel disease (IBD).@*METHODS@#Caco-2 cells were treated with 20% fibroblast conditioned medium or co-cultured with fibroblasts highly expressing WNT2b, with the cells without treatment with the conditioned medium and cells co-cultured with wild-type fibroblasts as the control groups. The changes in barrier permeability of Caco-2 cells were assessed by measuring transmembrane resistance and Lucifer Yellow permeability. In Caco-2 cells co-cultured with WNT2b-overexpressing or control intestinal fibroblasts, nuclear entry of β-catenin was detected with immunofluorescence assay, and the expressions of tight junction proteins ZO-1 and E-cadherin were detected with Western blotting. In a C57 mouse model of dextran sulfate sodium (DSS)-induced IBD-like enteritis, the therapeutic effect of intraperitoneal injection of salinomycin (5 mg/kg, an inhibitor of WNT/β-catenin signaling pathway) was evaluated by observing the changes in intestinal inflammation and detecting the expressions of tight junction proteins.@*RESULTS@#In the coculture system, WNT2b overexpression in the fibroblasts significantly promoted nuclear entry of β-catenin (P < 0.01) and decreased the expressions of tight junction proteins in Caco-2 cells; knockdown of FZD4 expression in Caco-2 cells obviously reversed this effect. In DSS-treated mice, salinomycin treatment significantly reduced intestinal inflammation and increased the expressions of tight junction proteins in the intestinal mucosa.@*CONCLUSION@#Intestinal fibroblasts overexpressing WNT2b causes impairment of intestinal mucosal barrier function and can be a potential target for treatment of IBD.

Levodopa combined with visual stimulation training in the treatment of ametropic amblyopia in adolescents / 国际眼科杂志(Guoji Yanke Zazhi)

· AIM:To explore the effect of levodopa and visual stimulation training in the treatment of adolescent ametropic amblyopia.· METHODS:Sixty-four adolescents (68 eyes) with ametropic amblyopia who treated in our hospital from May 2013 to May 2016 were included into this study,and the patients were randomly divided into the observation group (n=32,33 eyes) and the control group (n=32,35 eyes).The patients in the control group received visual stimulation training therapy,and the patients in the observation group were given levodopa on the basis of the control group.After treated for 24wk,the clinical effects and changes of P-VEP and visual sensitivity before and after treatment were compared.· RESULTS:After treated for 24wk,the total effective rate of the observation group (91％) was significantly higher than that of the control group (71％),and the difference was statistically significant (P ＜ 0.05).Although the amplitude of P-VEP P100 in the control group increased and the latency was shortened,the effect of the observation group was more significant,and the difference was significant (P＜0.05).· CONCLUSIONS:Levodopa combined with visual stimulation training has significant effect in the treatment of adolescent ametropic amblyopia,it can better correct the visual sensitivity,and improve optic nerve conduction function.

Clinical research on application of adjustable sutures in glaucoma filtering operation / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To discuss the application effect of adjustable sutures in glaucoma filtering operation after trabecular resection. METHODS: Seventy-eight cases ( 101 eyes ) suffered from glaucoma were randomly divided into two groups, observation group and control group. Thirty-nine cases ( 51 eyes ) in the observation group underwent trabeculectomy with adjustable sutures, the control group (39 cases, 50 eyes) only adopted trabeculectomy.RESULTS: Compared Preoperative IOP in two groups, the difference was not statistically significant (P>0. 05). After 6mo, IOP were decreased compared with preoperative in two groups, and that in observation group was lower than control group, the difference was statistically significant ( P CONCLUSION: The adjustable sutures combined with trabeculectomy for glaucoma can significantly reduce the postoperative complications. The curative effect is exact and clinically applicable.

Effects of short-term forest bathing on human health in a broad-leaved evergreen forest in Zhejiang Province, China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the effects of short-term forest bathing on human health.</p><p><b>METHODS</b>Twenty healthy male university students participated as subjects and were randomly divided into two groups of 10. One group was sent on a two-night trip to a broad-leaved evergreen forest, and the other was sent to a city area. Serum cytokine levels reflecting inflammatory and stress response, indicators reflecting oxidative stress, the distribution of leukocyte subsets, and plasma endothelin-1 (ET-1) concentrations were measured before and after the experiment to evaluate the positive health effects of forest environments. A profile of mood states (POMS) evaluation was used to assess changes in mood states.</p><p><b>RESULTS</b>No significant differences in the baseline values of the indicators were observed between the two groups before the experiment. Subjects exposed to the forest environment showed reduced oxidative stress and pro-inflammatory level, as evidenced by decreased malondialdehyde, interleukin-6, and tumor necrosis factor a levels compared with the urban group. Serum cortisol levels were also lower than in the urban group. Notably, the concentration of plasma ET-1 was much lower in subjects exposed to the forest environment. The POMS evaluation showed that after exposure to the forest environment, subjects had lower scores in the negative subscales, and the score for vigor was increased.</p><p><b>CONCLUSION</b>Forest bathing is beneficial to human health, perhaps through preventive effects related to several pathological factors.</p>