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.
Humans ; Mice ; Animals ; Caco-2 Cells ; beta Catenin/metabolism* ; Culture Media, Conditioned/pharmacology* ; Tight Junctions/metabolism* ; Intestinal Mucosa ; Inflammatory Bowel Diseases ; Tight Junction Proteins/metabolism* ; Inflammation/metabolism* ; Fibroblasts/metabolism* ; Mice, Inbred C57BL ; Glycoproteins/metabolism* ; Wnt Proteins/pharmacology* ; Frizzled Receptors/metabolism*

Neuropathic pain(NP) has similar phenotypes but different sequential neuroinflammatory mechanisms in the pathological process. It is of great significance to inhibit the initiation of neuroinflammation, which has become a new direction of NP treatment and drug development in recent years. Mongolian drug Naru-3 is clinically effective in the treatment of trigeminal neuralgia, sciatica, and other NPs in a short time, but its pharmacodynamic characteristics and mechanism of analgesia are still unclear. In this study, a spinal nerve ligation(SNL) model simulating clinical peripheral nerve injury was established and the efficacy and mechanism of Naru-3 in the treatment of NPs was discussed by means of behavioral detection, side effect evaluation, network analysis, and experimental verification. Pharmacodynamic results showed that Naru-3 increased the basic pain sensitivity threshold(mechanical hyperalgesia and thermal radiation hyperalgesia) in the initiation of SNL in animals and relieved spontaneous pain, however, there was no significant effect on the basic pain sensitivity threshold and motor coordination function of normal animals under physiological and pathological conditions. Meanwhile, the results of primary screening of target tissues showed that Naru-3 inhibited the second phase of injury-induced nociceptive response of formalin test in mice and reduced the expression of inflammatory factors in the spinal cord. Network analysis discovered that Naru-3 had synergy in the treatment of NP, and its mechanism was associated with core targets such as matrix metalloproteinase-9(MMP9) and interleukin-1β(IL-1β). The experiment further took the dorsal root ganglion(DRG) and the stage of patho-logical spinal cord as the research objects, focusing on the core targets of inducing microglial neuroinflammation. By means of Western blot, immunofluorescence, agonists, antagonists, behavior, etc., the mechanism of Naru-3 in exerting NP analgesia may be related to the negative regulation of the MMP9/IL-1β signaling pathway-mediated microglia p38/IL-1β inflammatory loop in the activation phase. The relevant research enriches the biological connotation of Naru-3 in the treatment of NP and provides references for clinical rational drug use.
Rats ; Mice ; Animals ; Matrix Metalloproteinase 9/metabolism* ; Rats, Sprague-Dawley ; Neuroinflammatory Diseases ; Interleukin-1beta/metabolism* ; Spinal Cord/metabolism* ; Signal Transduction ; Hyperalgesia/metabolism* ; Neuralgia/metabolism*

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.
Child ; Male ; Humans ; Female ; Child, Preschool ; Adolescent ; Crohn Disease ; Inflammatory Bowel Diseases ; Colon ; Inflammation ; Colonoscopy ; Glycoproteins ; Wnt Proteins

Obstructive sleep apnea (OSA) is a sleep disorder with a high incidence and severe impact on the human body, which can induce systemic chronic inflammatory responses. Chronic inflammation is an important cause of exacerbation of OSA and its associated complications. Nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) is an inflammasome that is widely found in epithelial cells and immune cells and plays an important role in inflammatory diseases as an important component of innate immunity. Research evidence suggests that the activation of NLRP3 inflammasomes can exacerbate the damage to neurons, endothelial cells, lung and kidney caused by OSA, and these effects can be eliminated by genetic or pharmacological deletion of NLRP3. Targeting inhibition of NLRP3 inflammasome may serve as a co-therapeutic strategy for OSA-induced related complications. This article reviews NLRP3 inflammasome and its mechanism in OSA-related concurrent diseases, which can provide scientific basis for prevention and intervention of OSA and its related complications.
Humans ; Inflammasomes ; Endothelial Cells ; NLR Family, Pyrin Domain-Containing 3 Protein ; Inflammation ; Sleep Apnea, Obstructive ; Nucleotides

Obstructive sleep apnea (OSA) is a sleep disorder with a high incidence and severe impact on the human body, which can induce systemic chronic inflammatory responses. Chronic inflammation is an important cause of exacerbation of OSA and its associated complications. Nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) is an inflammasome that is widely found in epithelial cells and immune cells and plays an important role in inflammatory diseases as an important component of innate immunity. Research evidence suggests that the activation of NLRP3 inflammasomes can exacerbate the damage to neurons, endothelial cells, lung and kidney caused by OSA, and these effects can be eliminated by genetic or pharmacological deletion of NLRP3. Targeting inhibition of NLRP3 inflammasome may serve as a co-therapeutic strategy for OSA-induced related complications. This article reviews NLRP3 inflammasome and its mechanism in OSA-related concurrent diseases, which can provide scientific basis for prevention and intervention of OSA and its related complications.
Humans ; Inflammasomes ; Endothelial Cells ; NLR Family, Pyrin Domain-Containing 3 Protein ; Inflammation ; Sleep Apnea, Obstructive ; Nucleotides

Objective To investigate the relationship between body composition and serum lipids and uric acid among adults in Maonan, and to analyze the effect of body composition changes on blood lipid and uric acid. Methods Totally 584 Maonan adult volunteers in Maonan village of Maonan Autonomous County in Guangxi, the age from 20 to 80 were recruited. The height was measured by the personal height tester; the body composition was measured by the ANITAMC-180 instrument; and the blood lipids and blood uric acid were measured by the Hitachi 7600 instrument. The obtained data were statistically analyzed by SPSS 20. 0. Results The age,height, weight, free fat mass, muscle mass, presumptive bone mass, body water, proptein,extracellular water, intracellular water, and waist-to-hip ratio were greater in Maonan men than in women (P<0. 05). However, whereas male fat content, body fat rate, and subcutaneous fat content were smaller than those of female (P < 0. 01). The total prevalence of hyperuricemia and hyperlipidemia in Maonan nationality was 13. 9% and 28. 4%, respectively. The prevalence of hyperuricemia and hyperlipidemia in males was higher than in females. In males, the body mass, body mass index, free fat mass, fat mass, muscle mass, presumptive bone mass, protein, extracellular water, body fat rate, visceral fat content, subcutaneous fat content and waist-hip ratio of the hyperlipidemia group were higher than the normal group (P<0. 05); and in females, the age, body mass index, fat mass, body fat rate, visceral fat content and waist-hip ratio of the hyperlipidemia group were higher than the normal group. In male, The body mass, free fat mass, presumptive bone mass, body water, extracellular water of the hyperuricemia group were higher than the normal group (P<0. 05); In females, the age, body mass, body mass index, fat mass, extracellular water, body fat ratio, muscle mass, visceral fat content, subcutaneous fat content, and waist-hip ratio of the hyperuricemia group were higher than the normal group. Conclusion The detection rate of hyperlipidemia and hyperuricemia in males of Guangxi Maonan nationality is all higher than that in females. The body composition is significant differences between the normal adults and the patients with hyperlipidemia and hyperuricemia of Maonan nationality in Guangxi.

The Lewis lung carcinoma (LLC) metastatic mouse model was used to investigate the effects of gefitinib and Sijunzi Tang (SJZ) on pre-metastatic niche. The experimental protocol was approved by the Ethics Committee which belongs to Cancer Hospital, Chinese Academy of Medical Sciences. To generate spontaneous lung metastatic models, 1×10⁶ luciferase-labeled LLC cells were injected subcutaneously in the shaved right flank of mice. One day after LLC inoculation, the mice were randomly divided into model (saline), gefitinib (50 mg·kg^-1) treatment, SJZ treatment (25.74 g·kg^-1), and co-treatment gefitinib with SJZ groups, with intragastrical administration. After 14 days of continuous administration, tumor size was detected by IVIS^® Spectrum system. The number of monocytes and neutrophils and the expression levels of chemokine receptors (CXCR1, CCR2) and carcinogenic gene (c-Kit), in peripheral blood, spleen and lung tissues of mice were determined by flow cytometry. The contents of interleukin-IL-1α (IL-1α) and interleukin-6 (IL-6) were detected by the enzyme linked immunosorbent assay (ELISA). After 21 days of treatment, tumors were surgically removed, weighed and the tumor volume was measured with vernier caliper and the antitumor effect of co-administration was evaluated. After 45 days of administration, the survival of mice was recorded. The results of flow cytometry showed that the percentage of neutrophils in gefitinib group, SJZ group, and co-treatment group was significantly decreased in the lung tissue compared to the model group (P<0.05 or P<0.01), but there was no significant difference between three treatment groups (P>0.05). In the mouse peripheral blood and lung tissue, compared with the model group, the expression levels of CXCR1, CCR2 and c-Kit on the surface of neutrophils and monocytes in SJZ group and co-treatment group decreased or decreased significantly (P<0.01 or P<0.05). However, there was a significant increase in the expression level of c-Kit on the surface of monocytes (P<0.05). In the mouse spleen tissue, the expression levels of CXCR1, CCR2 and c-Kit in the gefitinib group increased significantly (P<0.05), while decreased significantly in SJZ or co-treatment group (P<0.05). The results of ELISA showed that the content of IL-1α in SJZ group decreased significantly in the plasma of the mice compared with the model group (P<0.01) and the content of IL-6 in co-treatment group decreased significantly (P<0.05). Compared with the gefitinib group, the content of IL-1 in the co-treatment group decreased significantly (P<0.05). In the tumor tissues of mice, compared with the model group, the content of IL-1α in the co-treatment group decreased significantly (P<0.05). Furthermore, the content of IL-1α in co-administrated group and IL-6 in SJZ or co-treatment group decreased significantly compared with the gefitinib group (P<0.05). After 21 days of continuous administration, the tumor inhibition rates of gefitinib group, SJZ group and co-administrated group were 45.7%, 38.4%, and 84.8%, respectively. After 45 days of administration, the survival rate of the model group was 0%, whereas the gefitinib, SJZ or co-treatment group has a survival rate of 40%, 60%, or 60%, respectively. In summary, our study illustrated that Sijunzi Tang could improve the anti-tumor effect of gefitinib by regulating pre-metastatic niche.

Objective To observe the effects of different health education patterns on chronic nonspecific low back pain (CNLBP). Methods From September, 2016 to April, 2017, 75 patients with CNLBP after rehabilitation were randomly divided into control group (n=45) and Back School group (n=30). The Back School group received group teaching including physiological and anatomy of lumbar spine, ergonomics, healthy posture and function exercise, once a week for four weeks. While the control group received the booklets including the same contents. They were self-assessed with Visual Analogue Scale (VAS), Oswestry Disability Index (ODI), and the 36-item Short Form (SF-36) before education, and one and three months of follow-up in clinics, or with call back services and Wechat. The recur-rence frequency was recorded. Results The score of VAS increased in both groups in the follow-up, but increased less in the Back School group (Z>2.645, P<0.01). The recurrence frequency was less in the Back School group (Z=-2.082, P<0.05), with more ODI score (Z=2.265, P<0.05) after three months of follow-up. The bodily pain score of SF-36 was more in the Back School group after one and three months of follow-up (t>2.273, P<0.05). Conclusion Health education with Back School may benefit to maintain the curative effects and function, reduce recur-rence, and improve the quality of life of CNLBP patients.

Objective To investigate the predictive value of the whole nodule size and solid component size of lung adenocarcinoma manifesting as subsolid nodule(SSN) in three different dimensions for pathologic grade.Methods We evaluated retrospectively preoperative chest HRCT data of 125 patients with 127 SSNs surgically resected and pathologically conformed lung adenocarcinomas.All specimens were divided into two groups: a total of 69 SSNs in group A, including 22 AIS and 47 MIA;a total of 58 SSNs in group B, only including IAC.Computer aided diagnosis software were used to measure the one dimension maximum diameter of solid component with lung window setting(1D-SCLW),two dimension maximum diameter of solid component with lung window setting(2D-SCLW),one dimension maximum diameter of solid component with mediastinal window setting(1D-SCMW),two dimension maximum diameter of solid component with mediastinal window setting(2D-SCMW),one dimension maximum diameter of whole nodule with lung window setting (1D-WNLW), two dimension maximum diameter of whole nodule with lung window setting (2D-WNLW), and volume of solid component with threshold of-300 HU (SCT) of all SSNs.Results 1D-SCLW, 2D-SCLW,1D-SCMW,2D-SCMW,1D-WNLW,2D-WNLW and SCT of the group B were significantly larger than those of the group A(P=0.000).ROC analyses indicated that the diagnostic efficiency of SCT for the pathologic grade was the highest among 7 CT features(AUC=0.887, sensitivity:81%,specificity:93%);The cut-off values of 1D-SCLW,2D-SCLW,1D-SCMW,2D-SCMW,1D-WNLW, 2D-WNLW and SCT were 17.50 mm,14.75 mm,9.50 mm,7.75 mm,0.50 mm,1.25 mm and 139.00 mm3.Multiple Logistic regression analysis revealed that SCT was the independent predictor of pathologic grade(OR=4.978,95%CI=1.430-17.331,P=0.012).SCT of 139.00 mm3 or greater was a significant indicator of IAC.Conclusion Among the whole nodule size and solid component size of SSN in three different dimensions on preoperative HRCT, SCT is found to be the independent predictor of pathologic grade, which may provide reference for surgery.

Objective To investigate the predictive value of whole nodule size and solid component size of pulmonary subsolid nodules (SSNs)with different window setting on preoperative HRCT for pathologic grade in lung adenocarcinoma.Methods We retrospectively evaluated preoperative chest HRCT and pathological data of 125 patients with 127 surgically resected lung adenocarcinoma manifesting as SSNs.All specimens were divided into two groups:a total of 69 SSNs in group A,including 22 adenocarcinomas in situ (AIS) and 47 minimally invasive adenocarcinoma (MIA);a total of 58 SSNs in group B,including invasive adenocarcinoma (IAC).Observer 1 used computer aided diagnosis software to measure the volume of whole nodule with lung window setting (WNLW),volume of solid component with lung window setting (SCLW),volume of solid component with mediastinal window setting (SCMW) and volume of solid component with threshold of-300 HU(SCT) of all SSNs.Observer 2 randomly selected 50 SSNs and repeated all the measurements.The interobserver agreement regarding quantitative measurements were evaluated by using intraclass correlation coefficient(ICC).The differences of all quantitative features between two groups were evaluated by Mann-Whitney U test.All the quantitative features were evaluated by using univariate logistic regression analysis,significant quantitative features identified by univariate logistic regression analysis were included in the multivariate logistic regression and independent predictors of pathological grade were obtained.Receiver operating characteristic analysis was conducted for the independent predictive factors that exhibited statistically significant differences in the multivariate logistic regression.Results The interobserver agreement regarding quantitative features were excellent (ICC＞ 0.75).The WNLW,SCLW,SCMW and SCT of group B were significantly larger than those of group A (P＜ 0.001).The univariate logistic regression analysis indicated that WNLW,SCLW,SCMW and SCT were significant (P＜0.001),the multivariate logistic regression analysis indicated that SCT was the independent predictive factor (OR=1.013,95％CI:1.006—1.020,P＜0.001).When SCT larger than 139.00 mm3,SSN was significantly associated with IACs (AUC=0.887,sensitivity=81％,specificity=93％).Conclusion SCT of SSNs on preoperative HRCT can be used to distinguish between AIS-MIA and IAC,which may provide information for choice of operation.